Respiration Physiology 109 (1997) 177 194

The elephants respiratory system: adaptations to gravitational stress

R.E. Brown a,c, J.P. Butler a, J.J. Godleski a, S.H. Loring a,b,*
a Physiology Program, School of Public Health, Har6ard Uni6ersity, Boston, MA 02115, USA Department of Anesthesia, DA 717, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA c Zoological Institute, Department of Zoomorphology, Go teborg Uni6ersity, Medicinaregatan 18, S -413 90 Go teborg, Sweden b

Received 13 May 1997; received in revised form 22 May 1997; accepted 22 May 1997

Abstract Elephants have had to adapt to gravitational stresses imposed on their very large respiratory structures. We describe some unusual features of the elephants respiratory system and speculate on their functional signicance. A distensible network of collagen bers lls the pleural space, loosely connects lung to chest wall but appears not to constrain lung-chest wall movements. Myriad spaces within the network and its rich supply of capillaries suggest effective local sources and sinks for pleural uid that may replace the gravity-dependent ows of smaller mammals. The lung is partitioned into : 1 cm3 parenchymal units by a system of thick, elastic septa that ramify throughout the lung from origins on the lungs elastic external capsule. Parenchymal units suspended upon the elastic septal system protect dependent alveoli from compression, thereby reducing the usual gravitational gradient of lung expansion. Intra-pulmonary airways are devoid of cartilage, instead they appear to derive resistance to collapse from tethering forces of the attached septa. 1997 Elsevier Science B.V. Keywords: Elephant; Lung; Respiration; Anatomy; Histology; Mechanics; Gravity; Pleural space

1. Introduction While gravitational forces are known to inuence the mechanics of respiration (see West and Matthews, 1972; Bryan et al., 1966; Milic-Emili et al., 1966), we do not know if gravity has
* Corresponding author. Tel.: + 1 617 667-3092; fax: + 1 617 667-1500; e-mail: slor@chest.bidmc.harvard.edu

inuenced the design of the respiratory system. The elephant (Elephas maximus and Loxodonta africana ), largest extant land animal and second only to the largest whales in body mass, offers the opportunity to address important questions concerning the effects of gravity on the design of the mammalian respiratory system. Unlike other mammals, elephants have been thought not to have an intrapleural space (Todd,

0034-5687/97/$17.00 1997 Elsevier Science B.V. All rights reserved. PII S 0 0 3 4 - 5 6 8 7 ( 9 7 ) 0 0 0 3 8 - 8

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1913; Earles, 1929; Bu gen, 1988). Descriptions of attachments between lung and chest wall in elephants (connections form in late fetal life, Earles, 1929) have been in the literature since 1682 (Moulin (1682) cited by Earles, 1929). Early speculations about the physiologic importance of these connections (Todd, 1913; Earles, 1929), without apparent challenge in recent times (e.g. see Short, 1962; von Beyer et al., 1990) revolve around the elephants need for forceful inspiratory efforts to inhale through the long nasal tubes within the trunk and aspirate (and elevate) water into their trunk for drinking. Both maneuvers could require large transthoracic and trans-diaphragmatic pressures. It has been asserted that such large trans-respiratory pressures could result in a spontaneous pneumothorax or intrapleural hemorrhage were it not for the connections between the lung, chest wall and diaphragm (Todd, 1913; Earles, 1929; Short, 1962; von Beyer et al., 1990). As we will show, these assumptions are not consistent with known respiratory mechanics. Here we describe morphologic features (gross to ultrastructural) of the elephants respiratory system. We suggest that the morphology of the elephants lung indicates that gravitational forces do indeed inuence the design of the mammalian respiratory system, at least in this large species. The highly distensible connections between chest wall and lung cannot prevent a pneumothorax, nor do they affect the movement of the lung across the surface of the chest wall during breathing. Rather, the loose pleural space connective tissue may be important in the regulation of the ow of pleural uid. In the elephant the delicate gas exchanging pulmonary parenchyma is compartmentalized into grape-like units, : 1 cm3, by an extensive system of thick, elastic septa that originate from the lungs external capsule and which ramify throughout the lung. We suggest that the parenchymal compartments are suspended by the elastic septa so that dependent areas of the lungs are not compressed by the lung above, and nondependent alveoli are protected from overexpansion, thus reducing the effects of gravity acting on the elephants tall lung.

2. Materials and methods

2.1. The animal

An 18 year old, 2140 kg, adult male African elephant (Loxodonta africana ) suffering from chronic, localized osteomyelitis, was euthanized with alfentanyl, xylazine and phenobarbital. Other than the osteomyelitis no pathologic ndings were encountered during the necropsy begun immediately after death.

2.2. Thoracic dissection and gross obser6ations

With the animal in right lateral recumbency, the left fore and hind limbs were removed and the abdomen opened and eviscerated without penetrating the thoracic cavity. The thorax was initially opened and inspected via an incision through the left, dorso-lateral aspect of the diaphragms central tendon and later through a 20 20 cm opening made by cutting sections from three ribs in the left caudal, dorso-lateral rib cage (Fig. 1B). Following this, the left chest wall with its attached segment of diaphragm was removed from rst to last rib, sternum to the dorsal costal arch. Assessment of the mobility and deformability of the pleural connective tissue were completed at this time. Representative specimens of central tendon, diaphragm muscle, intercostal muscle and costal periosteum were harvested with attached pleural connective tissue. Lungs were removed en bloc with attached trachea and heart. Lung tissue was sampled from all regions, deep to supercial, of both lungs. A primary bronchus and one of its branches was serially sampled from where it entered the lungs hilum until it reached a diameter of B 1.0 cm. A section of mid-trachea (intrathoracic) with ve complete rings was preserved in glutaraldehyde.

2.3. Tissue preparation 2.3.1. Fixation Tissue specimens were rinsed with 0.05 M phosphate buffer (pH 7.2) and xed for \ 24 h with

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glutaraldehyde (3.0%, E.M. Grade, Tousimis) in 0.05 M phosphate buffer (pH 7.2) at ambient temperature. The xative was replaced twice within the initial 4 h. Some tissue blocks were stored in glutaraldehyde for later sub-gross dissection. Tissues for electron microscopy were postxed in 1% osmium tetroxide in 0.05 M cacodylate buffer (pH 7.2). All tissues for microscopic examination were dehydrated in a graded ethanolic series (70% to absolute). The elephants lung tissues were compared with those of three domestic species: mature cattle and horse, both : 400 kg, and pig, : 55 kg. Tissue from healthy lungs was harvested at necropsy and immersed in 10% buffered neutral formalin.

2.3.2. Light microscopy Specimens were processed according to standard parafn techniques. Sections, 6 v m thick, stained with hematoxylin and eosin, Verhoeff-Van Giesons and Massons Trichrome were photographed with a Nikon photomicroscope. Montage reconstructions were accomplished using overlapping elds digitally photographed (10 ) with a Dage 725 CCD camera on a Leitz photomicroscope into a Zeiss IBAS image analysis system. Individual elds were reassembled using Adobe-Photoshop running on an Apple Macintosh, Quadra 605. 2.3.3. Transmission electron microscopy Specimens were embedded in Spurrs resin according to standard procedures. Thin sections, B 100 nm, stained with uranyl acetate and lead citrate were examined with a Phillips 300 electron microscope.

3. Results

3.1. Gross obser6ations 3.1.1. The animal The maximum external dimensions of the thorax (with the animal in lateral recumbency prior to death) were: 118 cm transverse and 178 cm ventro-dorsal. The trunk, from its distal tip to approximately the level of the palate, was 200 cm long. The nasal tubes within the trunk were highly resistant to collapse; an investigator (101 kg) supported by one knee (68 cm2 of contact) on the distal end of the trunk did not collapse the enclosed nasal tubes upon the researchers ngers inserted into the external nares. 3.1.2. Thoracic ca6ity A small (15 cm) initial incision through the diaphragms central tendon immediately resulted in a large pneumothorax (Fig. 1). Surrounding the pneumothorax cavity was a lustrous, smooth surface formed of pale-yellowish connective tissue. This grossly homogeneous, loose connective tissue we refer to as pleural space connective tissue (PSCT). Through the chest-wall window in the

Fig. 1. Diagram of a section through the thorax of an African elephant (Laxodonta africana ) in right lateral recumbency. (A) Prior to opening the chest the lung is apposed to the chest wall and diaphragm. Pleural space connective tissue in the closed chest would form a layer a few cm thick lling the pleural space. (B) Thorax was initially opened and inspected through a diaphragmatic incision, internal to which an open, tissue free space, i.e. pneumothorax, was observed. Prior to any further disturbance to the thorax a 20 20 cm window was made in the lateral rib cage. The large pneumothorax was surrounded by a smooth surface of aerated pleural space connective tissue that prevented us from seeing the surfaces or shape of the lung, diaphragm or chest wall.

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caudo-dorsal rib cage (Fig. 1B), the PSCT surrounded a spherical pneumothorax cavity more than twice the diameter of the chest wall window, beyond which no thoracic viscera were visable. The aerated PSCT covered and lled all the space between chest wall, fully collapsed lungs, diaphragm and pericardium. Note that the PSCT would have had a much smaller volume in the intact (closed) chest before it became aerated by the pneumothorax and resulting lung collapse (compare Fig. 1A and 1B). When not stretched to its elastic limit, the PSCT was soft and could be easily deformed. However, when stretched as it was surrounding the pneumothorax cavity it was extremely tough and could not be penetrated by a nger. Handling and dissecting the unstressed PSCT was difcult, due to its slickness and the ease with which its substance was sheared when attempting to penetrate it. Examination of the lustrous PSCT attached to excised lungs and chest wall revealed it to contain myriad small spaces (uid and air lled) among the wet, shiny bers; the gross appearance was somewhat similar to a pad of wet, tightly woven, multi-layered surgical gauze. The PSCT appeared grossly to be structurally isotropic when aerated (two dimensionally isotropic in the tangent plane when not aerated) and without any obvious macro structure.

Fig. 2. Diagram of a 25 25 cm slab of the elephant diaphragms central tendon lying on a surface with the attached pleural space connective tissue uppermost. The surpercial tissue layer, grasped at one edge of the slab, could be displaced (shear deformation) more than 17 cm without apparently distrubing the next deeper layer.

3.1.3. Diaphragm The diaphragms insertion extended from the ventral end of the rst pair of ribs caudo-dorsally to the most dorsal aspect of the 21st (last) rib. The muscular portion of the diaphragm at mid-lateral thorax was approximately 3 cm thick and 17 cm long from rib cage to central tendon. 3.1.4. Deformability and mobility of pleural space connecti6e tissue We assessed the degree to which the PSCT might limit the mobility of the structures to which it was attached by the following: 1. Although the slimy PSCT was attached to and appeared continuous between the rib cage and the thoracic surface of the diaphragm muscle it did not impede the diaphragm from being elevated perpendicular to the inner surface of

the rib cage. With the diaphragm held perpendicular to the rib cage, the attached PSCT was not taut, but remained loose and easily deformed. 2. A 3 3 cm section of the central edge of the diaphragm muscle was isolated from surrounding muscle, but not from the underlying PSCT loosely binding it to the inner rib cage. This piece of muscle could be slid easily back and forth, away from and towards the diaphragms insertion on the rib cage, over a distance of 26 cm. 3. A slab of the diaphragms central tendon, 25 25 cm, was isolated without displacing the attached PSCT and placed with its abdominal side down (Fig. 2). At the midpoint along one edge of the slab, the most supercial (closest to lung in vivo) portion of the PSCT was grasped with forceps. The point of the forceps grasping the PSCT could be easily moved towards the opposite side of the slab, creating a deep V shaped pattern in the supercial PSCT, for a distance of \ 17 cm without dislocating or disturbing the immediately deeper layers of PSCT. The PSCT, although easily distended or deformed, was not elastic; for example, when the supercial layer was released it remained in its distorted V conguration, retracting towards its original position by less than 2 cm.

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Fig. 3. Elephant lung, supercial relationships of macroscopic connective tissue components, light microscopy reconstruction, lung xed at zero stress. The lungs highly distensible external capsule (Ext C) is folded or pleated when under no tension. The pleural space connective tissue (PSCT) is attached evenly across the lungs external capsule. The distensible intralung septa (ILS) originate from the lungs external capsule and ramify throughout the pulmonary parenchyma, dividing the parenchyma into compartments B 1 cm3.

After removing the lungs from the thorax, PSCT attached to the lung could be easily stretched to considerable lengths. A handful of PSCT lying next to the lung in the unstressed state could be easily stretched uniaxially to 20 50 cm without disturbing (elevating) the surface of the lung itself. The elongated stem of PSCT raised from the lung surface, when released, did not reform itself into the smooth layer covering the lung, but instead remained in a deformed, elongated blob lying on the surface. The PSCT was a very slimy material that stuck to most things it touched (e.g. latex gloves, table tops) but was difcult to grasp because it was so easily sheared (deformed). A layer with an unstressed (but aerated) thickness of \ 5 cm could be easily squeezed between thumb and nger to a layer B 1 mm thick.

length and when released returned to their original highly folded conguration (Fig. 3). From the external capsule strong, elastic connective tissue septa ramied throughout the pulmonary parenchyma. All grossly visible intrapulmonary airways were invested with a thick coat of dense connective tissue. The airways with their peribronchial connective tissue gave the cut surface of the lung a very rough, pebbly feel and appeared as if the walls of all airways contained abundant cartilage, although no cartilage was found on subsequent microscopic examination. When the cut ends of small, : 2.0 mm luminal diameter airways were grasped with forceps considerable force was needed to collapse the lumen. (This high resistance to the collapse of small airways is not a feature of the lungs of pig, cow, horse, human or most other terrestrial animals.)

3.1.5. Lungs The left and right lungs having one lobe each were encapsulated with a thick and highly elastic layer of dense connective tissue (Figs. 3, 4 and 6A). The PSCT completely covered and was attached to this capsule, which is homologous with the visceral pleura (albeit without a serosa, see below) covering the lungs of other animals. Isolated strips of the lungs thick, elastic capsule could be stretched greater than twice their original

3.1.6. Trachea The lumen of the mid-trachea, 7.5 5.5 cm, was supported by massive cartilaginous rings (Fig. 5). The end of each incomplete ring articulated with its other end and the opposite end of one of the adjacent rings. This produced a helical arrangement of the tracheal rings, the overlapped, alternating attachments making a saw-tooth pattern (zipper like) along the tracheas dorsal surface. The dorsal third (both sides) of each ring

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Fig. 4. Elephant lung \ 15 cm deep from its pleural surface showing macroscopic relationships of connective tissue components, light microscopy reconstruction, xed under zero stress. The distensible intralung septa (ILS) ramify extensively throughout the lung, dividing the pulmonary parenchyma into compartments B 1 cm3. The intralung septa when under no tension, as in this reconstruction, are folded and pleated. Note that the vessels (V) and bronchi (B) with luminal diameters B 0.1 mm are attached within and surrounded by the supporting framework of the intralung septa. When this gure and Fig. 3 are compared, it may be noted that the intralung septa occur in a range of thicknesses from that exceeding that of the lungs external capsule down to that only somewhat thicker than an alveolar septum.

overlapped substantially with the two adjacent rings. Thus, the borders of the rings were not parallel. The rings cross-sectional shape (in planes passing through the airway axis) varied from a thick oval with a convex external surface ventrally to that of a attened oval cross section laterally, ending with irregular facets for their double articulation dorsally. The rings were xed to one another in the overlapping areas and across their articulations by tough brous tissue. The tracheal mucosa had many 1 mm deep longitudinal rugae. These rugae could conceivably result from constriction of the trachea by contraction of the very thin (relative to that of 500 kg cattle) band of trachealis muscle, 1.5 cm long, which was inserted on the unyielding luminal surface of the rings, spanning their articulated ends. However, the brous tissue xing the rings dorsally and the thinness of the trachealis muscle make this seem unlikely, and we believe that the tracheal conformation we saw postmortem existed in vivo. The cartilaginous rings supporting the

trachea and extrapulmonary primary bronchi ended at the lung hilum.

3.2. Subgross dissection of lung tissue blocks

Blocks of lung, 2 1 1 cm, xed in glutaraldehyde were dissected and microscopically examined at 1040 . The xed PSCT, though less distensible than when fresh, could be easily deformed and elongated to more than twice its xed-resting length. The PSCT appeared as a complex, 3-D brous network containing myriad air- and uid-lled spaces B 0.1 mm3; the spaces between bers occupied a larger volume than the formed elements. In most areas the collapsed lungs external capsule was nely folded or pleated (Fig. 3). The connective tissue septa seen on gross dissection partitioned the pulmonary parenchyma into isolated compartments of about 1.0 cm3 (Figs. 3, 4 and 6D). We term these septa intralung septa to distinguish them from the alveolar septa compris-

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ing the parenchyma. The xed parenchyma could be easily scraped from the septa leaving strongwalled cavities. Commonly the thicker intralung septa could be easily separated into two distinct, tough layers between which were found scattered, loose collagenous attachments. Airways and vessels within the lung were invested with dense connective tissue continuous with the septa (Figs. 3, 4 and 8A). Each parenchymal compartment was apparently supplied by a single airway and vascular branch. These compartmental branches diverged from their parental airway or vessel (located within an adjacent intralung septum) and

coursed to the center of the parenchymal compartment before giving off a cluster of terminal branches. These compartmental branches and their terminal ramications (to the limit of microscopic observation) were covered by a reection of intralung septal tissue (Fig. 8A).

3.3. Microscopic anatomy 3.3.1. PSCT Other than the surface of the pericardium in contact with the heart, no mesothelial (serosal) lining was found within the thoracic specimens, including samples of the lungs external capsule deep to the PSCT, the PSCT and the internal chest wall and diaphragm (Fig. 6A and 6C). However, we found a normal appearing serosa investing the abdominal surface of the diaphragm. The PSCT, including its attachments to the external lung capsule, diaphragm and internal surfaces of chest wall, was almost entirely collagenous with but an occasional elastin ber ( B 1 per 40 eld) found within its structure where it joined the intercostal membrane. The intercostal membrane was a bi-layered structure, each layer of which appeared similar in thickness and elastin content to the lungs external capsule. Whereas in most of the PSCT there was no apparent organization of the collagen bers of the PSCT (Fig. 6A and 6C), in some areas small bundles of collagen bers were found to join and diverge in a chicken wire like arrangement (Fig. 6B). An abundance of small vessels, capillaries and perhaps lymphatic ducts, were found throughout the PSCT (Fig. 6B and 6C). The number of these vessels appeared excessive for the almost negligible metabolic needs of this collagenous tissue. Dispite the much greater density of formed elements in the lung capsule relative to the pleural space, there were about 7-times as many vessels per eld in the loose PSCT, : 7.5 per eld, as in the lung capsule. Further, about 1/2 of the vessels found within the lung capsule itself were found within the 15% of the capsule nearest the PSCT. Examination of the dense, pure collagenous tissues of the diaphragms central tendon and costal periosteum showed B 0.3 vessel per eld.

Fig. 5. Segement of elephant trachea taken midway from larynx to carina. (A) Perspective of complete trachea, dorsal aspect foremost. (B) Mid-sagital section of ventral aspect. (C) Cross section with dorsal aspect at bottom. The cartilaginous rings supporting the lumen are overlapped by 25 + % of their width along the tracheas lateral aspect (see cut off rings, cross-hatched in (A). The overlap is reinforced with a dense brous aponeurosis (FA) connecting adjacent rings. Over the ventral aspect of the trachea the rings do not overlap (see Fig. B) but continue to be strongly united by the brous aponeurosis. With a saw- tooth-like appearance, the end of each cartilaginous ring articulates with (1) its opposite end, (2) the opposite end of the adjacent ring and (3) continues to be overlapped with its adjacent member. The interlocked ends of the cartilaginous rings, similar to an osseous articulation, are supported by tough brous tissue (FA in C). Note that the cross-sectional shape of the cartilagenous rings changes considerably as one moves around their circumference. The tracheal mucosa (M) is strongly pleated into numerous longitudinal rugae.

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Fig. 6. Elephant lung, light microscopy, Verhoeff-Van Giesons elastin stain, collagen red, elastin black, smooth muscle brown. (A) Filling the center of the gure is the dense and highly distensible external capsule of the elephants lung. The external capsule has an elastin (black) content of at least 35% relative to that of collagen (red). Across the top of the gure, external to the lungs external capsule, is the diffuse, unorganized pleural space connective tissue (ct) composed entirely of collagen. Note the absence of any serosal surface on the external surface of the elephants lung or within the pleural space connective tissue. The pulmonary parenchyma along the gures lower edge is divided by one intralung septum originating from the lungs external capsule. Compare the thickness of the elephant lungs external capsule in this gure with that of the large domestic animals visible in Fig. 8D. (B) The pleural space connective tissue, in some areas appeared to have some microscopic organization (see text for details). Many small vessels (identied by their endothelial cells) can be found throughout the pleural space connective tissue. (C) The collagenous but highly distensible pleural space connective tissue is less organized in this view than in B, contained far more capillaries and lymphatics that required for the metabolic demands of this brous tissue. The lungs external capsule crosses the lower, left corner of this gure. (D) The branching (three) point of an intralung septum surrounded by distorted and collapsed parenchyma. Note the high elastin (black) content of the distensible intralung septum.

3.3.2. Capsule and septal system of lung The lung was enclosed within a thick (mean 0.70 mm, range 0.25 1.75 mm) capsule composed of 3550% elastin and 50 65% collagen (surface area approximations) (Figs. 3 and 6AFig.

7). Intralung septa, with a brous arrangement similar to that of the lungs external capsule, were continuous with the brous components of the external capsule (Figs. 3, 4 and 6A and 6D). The thickness of intralung septa varied from those

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equal to the lungs external capule to those only a few times thicker than an alveolar membrane. The thickness of a septum appeared to be independent of its position; thick and thin septa could be found both at the lung surface attached to the external capsule and deep within the lung. The elephant was found to have 2 4 times the number of intralung septal surfaces per eld as did the domestic animals examined for this study, although it is important to remember that there was an unknown degree of lung collapse in all the examined animals. All pulmonary vessels larger than capillaries and all non-respiratory airways were enclosed within intralung septa (Figs. 3, 4 and 8AFig. 9). The brous tissue of a septum was continuous with the brous tissue forming the wall of the vessel or airway. The brous tissue layer surrounding airways and vessels contained copious amounts of elastin external to the muscularis (Fig. 8A). Larger vessels and airways were often located within the junction of two or more septa.

3.3.3. Airways Except for the initial 5 cm of the intrapulmonary primary bronchus around which isolated cartilaginous plates could be identied, the intra-

Fig. 7. Elephant intralung septa, transmission electron micrograph. The highly distensible intralung septa are composed of \ 35% ( \ 50% in this view) amorphous elastin bers (E) with the remainder being bundles of collagen bers (C) and the occasional broblast (upper left). Bar = 1 mm.

pulmonary airways were devoid of cartilaginous tissue. The latter was surprising considering the pebbly feel of the lungs cut surface and the fact that cartilage is a prominent intrapulmonary airway component in other species, including the horse, cow and pig. A circumferential sheath of dense connective tissue, continuous with and identical in appearance to the intralung septa, surrounded the larger airways ( ] 2 mm luminal dia) (Fig. 9). A layer of adipose tissue crossed by many radial collagenous bands (peribronchial tissue in Fig. 9) separated the airways septal sheath from its thick wall. Septa radiated from the sheath in the plane of the airways axis and continued into the pulmonary parenchyma. Smaller airways ( B 2 mm luminal dia) were always tightly invested within a septum (Figs. 3, 4 and 8A). The brous tissues of the airway and associated septum combined to create thick airway walls whose thickness external to the mucosa was 30200% that of the luminal diameter. By contrast, terminal bronchioles leading to alveolar ducts had wall thicknesses a small fraction of the luminal diameter, similar to the airways from the other taxa examined for this study (Fig. 8B). The mucosa of all airways larger than terminal bronchioles was folded creating long, narrow, axially orientated crypts (Fig. 8A,Fig. 9) that mimic the mucosal rosetts found in lungs that are in an actively broncho-constricted state. We estimate that the surface created by the pleated mucosa was sufcient to smoothly cover a tube 2 \ 5 times the observed diameter of that airway. The mucosa contained an abundance of goblet cells (Fig. 8A). A lm of amorphous material, assumed to be mucus, coated the mucosa in most airways and rarely polymorphonuclear cell were found ( 4 1 per 40 eld). The submucosal and muscularis layers of the airway walls contained an abundance of elastin bers relative to that of the other taxa examined (Fig. 8A and 8BFig. 9). The orientation of the submucosal elastin bers was predominantly axial (mostly cross-sections of bers were found in airway cross-sections), with the elastin bers of the submuscularis and attached intralung septa oriented more nearly radially (longitudinal sections of bers found in cross-sections

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Fig. 8. Elephant lung, light microscopy, Verhoeff-Van Giesons elastin stain. (A) Smaller conducting bronchus with attached septum. The mucosa of the airways in these lungs xed at zero stress was always found highly pleated. The majority of the submucosal elastin bers (black) have an axial orientation, in contrast to the elastin bers of the attached intralung septum which are radial to the airway. sm, airway smooth muscle. (B) Terminal bronchiole and alveolar duct. Numerous, axially oriented elastin bers lying deep to the airways mucosa (arrowhead) continue into the alveolar duct (top 3/4 of gure). The pulmonary parenchyma is highly distorted and collapsed in these lungs xed at zero stress (relaxed). Note the magnication of this gure is twice that of all other color micrographs. (C) These three micrographs represent one complete and continuous slice through the diaphragms central tendon from abdominal (1) to thoracic (4) sides (about 10% of the central tendons thickness is not represented in these three micrographs). The central tendon, \ 3.0 mm thick, was composed of three distinct and separate layers only loosely joined by occasional collagen bers (see left side of 3-4). The layer on the abdominal side of the central tendon (1-2), invested with a serosal membrane (not shown), was the only layer to contain elastin bers. Pleural space connective tissue was attached to 4 side of section 3-4. (D) External lung capsule of cow (D1), pig (D2) and horse (D3). Compare these with the elephants much thicker and denser external lung capsule seen in Fig. 6A. In addition, the elephants external capsule contained much more elastin. All micrographs of external lung capsule, e.g. Fig. 6A and D of this gure were stained simultaneously and are shown at identical nal magnication.

of airways). The submucosa of terminal bronchi contained abundant, axially-oriented elastin bers that continued into the alveolar ducts (Fig. 8B).

3.3.4. Gas exchange tissues The lungs were collapsed at the time of initial examination and during xation; thus the ne alveolar architecture was distorted or obscured

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Fig. 9. Intralung septa tethering a large airway. All airways had one or more intralung septa (ILS) attached to, and radiating normal to the airways axis. In the larger airways, such as the one shown here, the multiple septa were attached to the airway via a circumferential septum that enclosed a zone of interwoven collagenous bers and adipose tissue (peribronchial tissue). The submucosal layer contained large numbers of predominantly axially oriented elastin bers, seen here as a thick dense-black band deep to the airways pleated mucosa (M).

(Fig. 6DFig. 8BFig. 10). Many collagen bers and bundles of collagen bers were found immediately beneath the pulmonary epithelium and deeper within the alveolar membranes (Fig. 10). The apparently increased collagen content notwithstanding, the elephants alveolar membranes, type 1 and 2 pulmonary epithelial cells and capillaries, appeared like those of other mammals. Gas exchange tissue lined the intralung septa.

contained some elastin bers mingled among the collagen bers (Fig. 8C). Electron microscopy revealed a highly anisotropic arrangement of the collagen bers within each of the layers, and light micrographs suggested a different orientation of the bers in the different layers. The three layers were loosely joined by diffuse collagen bers.

3.4. Comparati6e morphology

In conjunction with the present study the lungs of domestic cow, horse, and pig were examined histologically (Fig. 8D). In these domestic animals, the lungs external brous capsule was always less than 0.18 mm thick (elephants capsule was 0.251.75mm thick) and only rarely were elastin bers identied (elephants capsule was elastin rich). The intralung septa of these animals were commonly composed of two thin collagenous sheets, which together were always less than 0.08 mm thick (not counting any gap between layers), and only very rarely contained elastin

3.3.5. Inammatory cells Alveolar macrophages were found within the elephants lung. Mononuclear cells found within alveolar capillaries have provisionally been identied as pulmonary intravascular macrophages (Fig. 10). 3.3.6. Diaphragm The central tendon over 3 mm thick was composed of three distinct layers of dense, collagenous tissue. The layer nearest the abdomen also

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bers. In contrast, the elephants elastin rich intralung septa were found in a range of thicknesses up to that of its external lung capsule. The interlung septa of the domestic taxa were only infrequently found to attach to an airway. In sharp contrast to those of the elephant, the larger intrapulmonary airways of these domestic animals contained cartilaginous rings while smaller, nonrespiratory airways contained cartilaginous plates. The thickness of the walls of smaller airways in these domestic animals was B 25% of the airways submucosal diameter (elephants airway walls were 30200% of submucosal diameter). Relative to those of the elephant, the wall of the airways of these domestic animals contained very few elastin bers.

respiratory system. The histologic ndings resulted from a random sampling of the lung so that a comprehensive morphometric evaluation of the regional differences in lung morphology was not possible.

4. Discussion

4.1. Mechanics of breathing and the elephants pleural space connecti6e tissue 4.1.1. Lung mo6ement relati6e to chest wall during breathing The mobility, extreme compliance, ease of shearing and slipperiness of the pleural space connective tissue (PSCT) implies that it has negligible inuence on the sliding motions of the lung relative to the chest wall during breathing. Thus, the lung is free to slide relative to the chest wall, diaphragm, mediastinum, and move in and out of the costophrenic sulcus. Also the fact that the lungs immediately collapse producing a large pneumothorax via a small incision through the diaphragm with an otherwise intact chest wall demonstrates, that the highly deformable PSCT does not restrict lung movement and cannot maintain the lungs in their inated conguration in the absence of an intact lung and chest wall, i.e. the pleural space must be closed for the elephant to breath. In that the elephants chest lacks both visceral and parietal mesothelial (serosal) surfaces, and its lung, albeit loosely, is attached to the chest wall, one could argue that the elephant lacks a pleural space. On the contrary, the elephant does possess a functional pleural space that allows independent movement between lung, chest wall and diaphragm, functioning identically to the liquidlined pleural space of other mammals. Unique to the elephants pleural space is that it is nely divided by the brous continuum of PSCT, the bers of which are lubricated by the elephants pleural uid. We consider the deformability or stretchiness of the PSCT to be a function of the network organization of its bers which are almost entirely inelastic collagen. While the ease with which it can

3.5. Critique of methods

Although we thoroughly examined the thoracic compartment and many areas within the lung, this study presents data on a single animal. In addition, except for examining the mobility of the PSCT, we have no in vivo or in situ experimental measurements of the mechanics of the elephants

Fig. 10. Elephants alveolar septum, transmission electron micrograph. Note the large quantity of collagen bers and bundles of collagen bers (arrowheads) lying between the pulmonary epithelium (PE) and capillary endothelium (CE). We have provisionally identied the large mononuclear cell lling the capillary lumen as a pulmonary intravascular macrophage. Bar = 1 mm.

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be sheared is, quite frankly, amazing, there are other examples (different taxa and different anatomical locations) in which networks of collagen bers undergo large physiologic deformations. A common example is the subcutaneous tissue attaching skin to underlying muscle fascia, e.g. the domestic cats thigh and knee can move cranio-caudally \ 15 cm beneath the skin of the ank without disturbing the overlying skin.

sphere); such pressures are remote from those anticipated during drinking in even the largest elephants. Thus, with a closed pleural space the elephants lungs do not need, nor do they have (see above) any attachments to the chest wall capable of preventing a pneumothorax while elevating water in the trunk or overcoming high resistances to inspiratory airow.

4.1.2. Ele6ating water 6ia the trunk during drinking Speculations in the literature suggest that the connections between the elephants lung and chest wall, i.e. PSCT are necessary to prevent a spontaneous pneumothorax and pleural vessel hemorrhage from occurring secondary to either elevating water in the trunk for drinking or forceful inhalations against a high resistance to ow through the long trunk (we consider a high resistance to airow doubtful but no measurements are available) (Todd, 1913; Earles, 1929; Short, 1962; von Beyer et al., 1990). Large trans-respiratory pressure differences (from alveolar space to body surface), up to 300 cmH20 in a large elephant completely lling its nasal tubes to the level of the palate, would be born by the chest wall and diaphragm as the elephant aspirated water up the trunk with its respiratory muscles. We assume (1) that elephants nearly ll their trunks with water and (2) that they use ventilatory muscles rather than lingual and facial muscles. Yet, the pressure difference between alveolar space and pleural space (transpulmonary) tending to distend pulmonary parenchyma is strictly a function of lung volume and is governed entirely by the elastic recoil properties of the lung per se. It follows that the elephants transpulmonary pressures are unaffected by drinking and thus the likelihood of a spontaneous pneumothorax (breech of pleural integrity) is not only no higher than that of other animals but probably much lower because of the elephant lungs very thick external capsule. Further, cavitation (i.e. separation of tissues due to extremely low pressure) within the elephants pleural space secondary to large transthoracic pressure differences cannot occur at absolute pressures above 50 torr ( 710 torr relative to atmo-

4.2. Mechanical implications of ele6ating water in a long trunk

However the extraordinarily low intrapulmonary air pressures, 300 cmH2O, required to elevate water to a height of 3 m in the nasal tubes with respiratory muscles could potentially (1) collapse the extrathoracic trachea or nasal tubes, (2) injure the diaphragm (muscle, central tendon) or chest wall or (3) cause blood shifts or other cardiovascular effects (not discussed).

4.2.1. Nasal tubes and trachea Water aspiration to heights up to 3 m is possible only if the extra-thoracic trachea and nasal tubes within the exible trunk can resist collapse from such large transmural pressure differences. The resistance to collapse of the muscular and connective tissue elements forming the trunk which has no ossied tissue are a function of the trunks thickness. An indication that the trunk is sufciently resistant to collapse is that a local compressive stress \ 6.0 103 cmH2O failed to collapse the distal nasal tubes where the trunkwall is thinnest. Collapse of the extra-thoracic trachea is prevented by the massive tracheal rings which are functionally complete by virture of their interlocked ends in addition to the overlap between adjacent rings. The intrathoracic trachea, like the lung, is not affected by the trans-respiratory pressures associated with elevating water in the trunk for drinking. 4.2.2. Diaphragm and chest wall Production and support of the trans-respiratory pressure differences necessary to elevate water to the top of the trunk is no mean feat and suggests the presence of special structural modications within the elephants chest wall, i.e. ribs, inter-

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costal muscles and diaphragm. While we have no information concerning the mechanical properties of the ribs and intercostal muscles (gross and histological examination of the intercostal muscles were unremarkable), there are several features of the elephants diaphragm that may assist in the production and support of those forces. The thick, well developed, trilaminar structure of the elephant diaphragms central tendon appears to be an adaptation for the support of large trans-diaphragmatic pressures (Fig. 8C). Sacks and Chuong (1992) describe the organization of the collagen bers of the single layered central tendon of other mammals as orthotropic (having parallel bers; see also Grifths et al., 1992). There is a weakness with such an arrangement in that a rupture or separation could occur between bers at forces far below those required to break the bers themselves. Whereas the collagen bers of each individual layer of the elephants central tendon are also in a parallel arrangement, the different alignments of the bers in the three layers of the elephants central tendon would confer nearly isotropic mechanical properties to this trilaminate structure. The elephants diaphragmatic muscle appears to be well adapted for the generation of large forces and thus pressures. Using approximations of the internal dimensions of the elephants chest (taken here to be lateral = 100 cm and ventrodorsal = 143 cm) we can calculate the oval circumference (388 cm) and axially projected area of the diaphragm (Areadi) to be 1.12 104 cm2. From the internal circumference of the chest wall and thickness of the diaphragmatic muscle (taken here to be 3 cm) we can calculate the total cross sectional area of the muscle (CSAM) to be 1.16 103 cm2. (This underestimates both projected area and muscle cross-sectional area because of the oblique orientation of the elephants diaphragm.) Assuming the elephants diaphragmatic muscle generates a maximal stress (s) (tension) similar to other animals skeletal muscles (2 106 dynes/ cm2; Powell et al., 1984), we calculate the maximal pressure (Pdi) that this elephants diaphragm can produce is

Pdi =

|M CSAM : 200 cmH2O Areadi

The orientation of the surface of the elephants diaphragm is oblique (nearly 45) to the spinal axis (and gravitation force), in contrast to the more perpendicularly disposed diaphragms of other animals. This may represent an adaptation for the appropriate generation of pressures involved in drinking, or an adaptation for the support of the static pressure gradients associated with gravity acting on the very different densities of thoracic and abdominal viscera. These possibilities remain uncertain and deserve further study.

4.3. The elephants respiratory system: adaptations to gra6itational stress

The question, Do gravitational forces set limits on the design of the respiratory system? (Leith, 1976), has been long standing in respiratory physiology. At least two features of the elephants respiratory system appear to be structural adaptations for the amelioration of the effects of gravity: (1) capsule and intralung septal system and (2) PSCT.

4.3.1. Capsule and intralung septal system of lung The total pressure difference (P ) from the top to the bottom of the lung resulting from gravity and tending to collapse dependent alveoli and hyperexpand superior parenchymal tissue increases directly with the height of the lung: P = pgh, where p, tissue density, g, acceleration due to gravity and h, height of lung. West and Matthews (1972) described that in most mammals there is a regional (top to bottom) difference in alveolar ination existing at low lung volumes resulting from the compressive effects of gravity, but that as lung volume is increased the regional differences would disappear due to a stiffening effect due to the parenchymas nonlinear stress-strain characteristic (see Radford, 1957). That is, as the alveoli in the superior areas of the lung reach their maximum volume their stiffened parenchymal components force further ination to occur in the more dependent and less distended regions of the lung.

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In the elephant, the pressures inating the alveoli would vary by 50 cmH2O from the top to the bottom of the lung estimated to be 145 cm tall at its maximum (in vivo density of lung taken to be z = 0.35 g m/cm3; see Ganesan et al., 1995). Assuming the elephants lung has a pressure-volume relationship similar to that found in other mammals in which the vital capacity spans a transpulmonary pressure range of 25 30 cmH2O, the 50 cmH2O pressure difference from top to bottom of its tall lung would mean that there would be no range of lung volumes at which both uppermost and lowermost alveoli would experience physiologically normal distending pressures during breathing. It follows that at all physiologic lung volumes, the elephants tall lung, without structural modications, would have a large proportion of its dependent alveoli collapsed and a large proportion of its superior alveoli over-distended or highly stressed. Here, it is important to distinguish between the vertical gradient in pleural pressure, Ppl, which supports the whole lung in gravity, and the vertical gradient in distending pressure of the pulmonary parenchyma, which we argue for the elephant must be substantially less than Ppl. In the lungs of other (smaller) mammals it is argued that the pressure distending the parenchyma is the elastic recoil pressure of the lung, Pel( ) = PA Ppl L (Mead et al., 1970) where PA is alveolar pressure. This pressure is also the stress within the parenchyma averaged over an area transecting many alveoli (e.g. several mm2). In the elephant this assumption may not hold; within the small parenchymal units stresses averaged over several mm2 may be substantially less than the stresses within the lung and its supporting intra-lung septal system averaged over many cm2. At least in very large lungs it appears unreasonable to expect lung tissue to be self supporting. If the mass of parenchyma, blood supply and airways comprising the elephants lung were to be supported solely from ne internal structures at the alveolar and ductal level it seems reasonable to expect considerable reinforcement within the lung. In fact, elements capable of supporting compressive stress would be necessary within its dependent portions. Such a structural adaptation at the level of the

alveolar membrane has not been found, or not recognized, in large mammals (including the elephant). We suggest that the elephant lungs capsule and intralung septal system is a structural mechanism that supports parenchymal, vascular and airway structures so that the alveoli are functionally isolated (partially or totally) from the gravitation effects described above. The thick external capsule (visceral pleura) of the elephants lung, apposed to the chest wall and diaphragm by virtue of a closed pleural space as in other mamals, serves as a foundation upon which to anchor the intralung septa (Figs. 3, 4 and 6A and 6D). The extensive ramication of the intralung septa throughout the pulmonary parenchyma serves as a space lling, interconnected set of surfaces tessellating the lung and supporting the resulting small ( B 1 cm3) mechanically isolated compartments of parenchyma. The high elastin content and coextensive network of collagen and elastin bers found in the elephant lungs intralung septa (and external capsule) (Fig. 6A and 6DFig. 7) is very similar to that found in mammalian Lig. nuchae and the birds Lig. propatagiale. Those highly distensible, elastic ligaments have long, linear segments of their force-length curves which enable physiologic changes in strain to be accompanied by very small changes in stress (Brown et al., 1994). In sharp contrast, pulmonary parenchymal strips have been shown to stiffen rapidly with very small increases in strain within their physiologic range (Radford, 1957). The collagen and elastin composite forming the structural elements of the elephant lungs external capsule and intralung septal system would allow these structures to maintain nearly constant support of the pulmonary parenchyma across a wide range of lung volumes. Furthermore, such pre-stress of the highly elastic tissue of the septa need not substantially reduce the compliance of the lung during volume expansion. That is to say, if pre-stress of the septal system at low lung volume were such as to suspend parenchymal compartments uniformly, there would be little regional difference in the ination of the elephants alveoli over a wide range of lung volumes. Indication that prestress conditions do exist within the elephant lungs highly elastic ex-

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ternal capsule and intralung septa is reected in their tightly folded (pleated) conformation seen in the deated, relatively gas free lung and in isolated tissue blocks. The linear elastic support mechanism provided over the entire lung by the elephant lungs capsule and intralung septal system is quite different from the strongly nonlinear properties ascribed to the self- supporting parenchymal tissue of other mammals (see Radford, 1957; Mead, 1961; Mead et al., 1970; West and Matthews, 1972). We would expect the compliance of the elephants pulmonary parenchyma within a single parenchymal compartment to be similar to that of other mammals and its total lung compliance (governed by the linear elastic force-length properties of septal system) to be similar to that of other mammals. That is, the elephants vital capacity could span a range of 2530 cmH2O as found in other mammals. However, if the elephant is to experience ventilation of all its alveoli, its lung-wide mean distending pressure (as well as Ppl) must be higher than other animals, at least 50 cmH2O. Over a 7-fold range of body mass there appeared to be no difference in external capsule or intralung septal thickness among the domestic animals. Additionally, the external capsule and intralung septa of the domestic animals was almost entirely collagenous. Yet between cattle and elephant (5-fold difference in body mass) there was more than a 3-fold difference in capsule thickness and an order of magnitude difference in average septal thickness (compare Fig. 6A, D, Fig. 8D). Additionally, the elephants lung has 2 4 times the density of intralung septa. Those results indicate that above a certain size, lung parenchymal tissue itself may not be self-supporting in the fashion described by Milic-Emili et al. (1966) and West and Matthews (1972). This transitional size appears to be one at which the difference in average recoil pressures between the top and bottom of the lung approaches the difference in recoil pressures between relatively un- inated and fully-inated parenchyma. The very sparse ( B 5%) elastin content of the intralung septa of the pig, cow and horse (and many other mammals, data not presented here), relative to that of the elephant (3550%), indicates that the pul-

monary mechanics of those species is governed by a parenchymal force-length relationship similar to that measured by Radford (1957); i.e. the nearly inextensible alveolar septa dominate overall lung recoil. In the domestic species the inelastic septa would function in parallel with the inelastic alveolar septa at high stresses.

4.3.2. Elastin content in walls of large airways and 6essels The walls and investing septa surrounding the elephants intrapulmonary airways and vessels contain many axially oriented elastin bers (Fig. 8A). If the extensibility of the elephants intrapulmonary airways and vessels is matched to that of the capsule and intralung septal system all these components would contribute to the mechanical support of the pulmonary parenchyma. While Hoppin et al. (1977) demonstrated that intrapulmonary mechanisms exist to isolate and protect the bronchial tree from axial distortion, we would suggest that the elephants elastic bronchial tree undergoes considerable length changes. Lai-Fook (1979) indicates that the larger arteries within the lung, function to limit regional expansion of pulmonary parenchyma, contributing to the pulmonary interdependence described by Mead (1961) and Mead et al. (1970). However, the dense elastic coat of the elephants pulmonary vasculature and the septal tissues investing it indicate that these vessels are quite extensible and that they probably lengthen as the surrounding tissue expands, contributing to local pressure-volume characteristics without limiting expansion. 4.3.3. Flow of pleural uid Pleural uid, under gravitational inuences, exhibits at most a hydraulic gradient from the top of the lung to the more dependent areas of the intrapleural space (Lai-Fook et al., 1984; Miserocchi et al., 1988). Departures of liquid pressure gradients from 1 cmH2O/cm vertical height have been associated with viscous pressure losses arising from pleural uid drainage (Lai-Fook and Rodarte, 1991) and may also be involved with pumping of pleural uid secondary to oscillating shear stresses during breathing (Butler et al., 1995). We suggest that the brous matrix of the

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PSCT (Fig. 6A, 6B and 6C) may be necessary to retard gravitationally driven ow of pleural uid from top to bottom of the elephants tall lung, which could result in excessive thinning of the lubricating layer of liquid between lung and chest wall and collections of pleural uid in the ventral thorax. The myriad small spaces within the brous continuum of the PSCT may act to slow the ow of pleural uid via a process of percolation through the brous matrix. Thus, the pleural uid would act as a lubricant between lung and chest wall by lubricating the motion between adjacent bers of the PSCT. The abundance of small vessels and lymphatics (Fig. 6B and 6C) throughout the PSCT suggest that large local uxes of pleural uid occur within the tissues brous matrix. Thus, in contrast to the pattern of pleural uid ow in other mammals with a uid lled pleural space, the elephants pleural uid may be produced and reabsorbed locally, reducing or eliminating the ux of uid from top to bottom of the thorax.

4.4. Maintenance of airway patency and the elephants intralung septa

During expiration, when the pressure driving ow is coupled to peribronchial pressure, ow limitation could potentially occur in any unsupported intra-thoracic airway. In contrast to other mammals, the elephant has no cartilaginous support of its intrapulmonary airways to maintain airway patency. However, the elephants intralung septa which surround and attach to airways larger than about 2 mm luminal dia. radiate normal to the airways axis and ultimately connect to the lungs external capsule, suggesting that the luminal patency of these larger airways may be maintained by a tethering mechanism (Fig. 9). Luminal patency of the smallest divisions of the bronchial tree, e.g. terminal bronchioles and alveolar ducts, in elephants (see below) as in other animals, is probably maintained by local radial tethering forces of the alveolar septa. However, the tethering open of the larger airways is by their attached intralung septa and is not dependent on local parenchymal tissue. The highly elastic intralung

septa, with a nearly linear force-length relationship in their range of physiologic lengths, could preserve airway patency at all lung volumes. This may represent an alternative adaptation for the airway luminal support provided by cartilage in other mammals. In the small respiratory and terminal bronchioles airway patency must be maintained solely by the local radial tethering forces of attached alveolar membranes in a fashion identical to that of other mammals (Fig. 8B). Many small bronchi, 5 2 mm luminal dia., which in other mammals would be supported by cartilaginous plates, had only one or two tethering intralung septa, so an alternative mechanism of luminal support may be operative in the smaller airways. The thick, but non-cartilaginous, walls of these small airways and their associated, investing septal tissues produce airways that are surprisingly resistant to collapse and that produce a pebbly texture to the cut surface of the lung. The bending stiffness of a material is a direct function of its thickness, so that the thick, dense walls of the smaller airways may protect their lumens from collapse. The relatively gas-free state of the intact lungs following their removal from the thorax was evidence that small airways remained patent at very low distending pressures. Interestingly, the folding pattern of the elephants airway mucosa, apparently resulting from reductions in airway diameter with the unopposed elastic recoil of the septal and capsular tissues (Fig. 8AFig. 9), appear identical to the mucosal rosetts that occur coincident with the bronchoconstriction produced by activation of airway smooth muscle. With reductions in airway caliber to the degree responsible for the observed mucosal folding, there would be substantial increases in airway wall thickness. It is possible that the elephants airway walls in vivo were no thicker than comparable airways in other animals. If that is the case it may be that the resistance to compression of the elephants smaller airways, as well as its larger airways, is conferred by the radial traction of the attached intralung septa. We suggest that the development of the highly elastic and extensive capsule and septa system of the elephant lung was driven by gravitational

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R.E. Brown et al. / Respiration Physiology 109 (1997) 177194 Afrikanischen Elefanten als Grundlage fu r tiera rztliches Handeln. Diss. Tiera rztliche Hochschule Hannover. Butler, J.P., Huang, J., Loring, S.H., Lai-Fook, S.J., Wang, P.M., Wilson, T.A., 1995. Model for a pump that drives circulation of pleural uid. J. Appl. Physiol. 78, 23 29. Earles, N.B., 1929. The anatomy of a foetal African elephant, Elaphus africanus (Laxodonta africana ), part 3, the contents of the thorax and abdomen and skelton. Trans. Roy. Sco. Edin. 56, 203 246. Ganesan, S., Rouch, K.E., Lai-Fook, S.J., 1995. A nite element analysis of the effects of the abdomen on regional lung expansion. Respir. Physiol. 99, 341 353. Grifths, R.I., Shadwick, R.E., Berger, P.J., 1992. Functional importance of a highly elastic ligament on the mammalian diaphragm. Proc. R. Soc. Lond. B. 249, 199 204. Hoppin, F.G. Jr., Hughes, J.M.B., Mead, J., 1977. Axial forces in the bronchial tree. J. Appl. Physiol. 42, 773 781. Lai-Fook, S.J., 1979. A continuum mechanics analysis of pulmonary vascular interdependence in isolated dog lobes. J. Appl. Physiol. 46, 419 429. Lai-Fook, S.J., Beck, K.C., Southorn, P.A., 1984. Pleural liquid pressure measured by micropipettes in rabbits. J. Appl. Physiol. 56, 1633 1639. Lai-Fook, S.J., Rodarte, J.R., 1991. Pleural pressure distribution and its relationship to lung volume and interstitial pressure. J. Appl. Physiol. 70, 967 978. Leith, D.E., 1976. Comparative mammalian respiratory mechanics. The Physiologist 19, 485 510. Mead, J., 1961. Mechanical properties of the lungs. Physiol. Rev. 41, 281 330. Mead, J., Takashima, T., Leith, D.E., 1970. Stress distribution in lungs: a model of pulmonary elasticity. J. Appl. Physiol. 28, 596 608. Milic-Emili, J., Henderson, J.A.M., Dolovich, M.B., Trop, D., Kaneko, E., 1966. Regional distribution of inspired gas in the lung. J. Appl. Physiol. 21, 749 759. Miserocchi, G., Kelly, S., Negrini, D., 1988. Pleural and extrapleural interstitial liquid pressure measured by cannulas and micropipettes. J. Appl. Physiol. 65, 555 562. Moulin, A., 1682. An Anatomical Account of the Elephant Burnt in Dublin. London. Powell, P.L., Roy, R.R., Kanim, P., Bello, M.A., Edgerton, R., 1984. Predictability of skeletal muscle tension from architectural determinations in guinea pig hindlimbs. J. Appl. Physiol. 57, 1715 1721. Radford, E.P., 1957. Recent studies of mechanical properties of mammalian lungs. In: Remington, J.W. (Ed.), Tissue Elasticity. Am. Physiol. Soc., Washington, D.C., pp. 177 190. Sacks, M.S., Chuong, C.J., 1992. Characterization of collagen ber architecture in the canine diaphragmatic central tendon. J. Biomech. Eng. 114, 183 190. Short, R.V., 1962. The peculiar lungs of the elphant. New Sci. 316, 569 571. Todd, T.W., 1913. Notes on the respiratory system of the elephant. Anat. Anz. Jena. 44, 175 183. West, J.B., Matthews, F.L., 1972. Stresses, strains, and surface pressures in the lung caused by its weight. J. Appl. Physiol. 32, 332 345.

demands for parenchymal support, but that with attachment to the airways, it subsequently replaced the need for cartilaginous support of airways lumens. The presence of cartilaginous rings supporting the lumens of the elephants extra-pulmonary bronchi and trachea demonstrates that elephants are indeed capable of developing such airway support structures.

Acknowledgements We thank Professor Kurt Benirschke for introducing us to this interesting problem, for his encouragement and for discussion of the ideas presented here. Discussions with Drs Fred Hoppin and Jere Mead led to some of the ideas presented here. We thank Bruce Ekstein, Tori Hatch and Bonnie Meek for their efforts in the preparation of the gures. We greatly appreciate the opportunity extended by the Wildlife Safari animal park, Roseberg, Oregon to attend the necropsy of their elephant. Dr Jack Mortenson and Amanda Sallon of Wildlife Safari and the necropsy team from the Oregon State University, College of Veterinary Medicine led by Dr Olaf Hedstrom were most supportive of our efforts. Dr George Kennedy of the College of Veterinary Medicine, Kansas State University generously supplied the domestic animal lung tissues. This work was supported by the Beth Israel Anesthesia Foundation and HL 52586.

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