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Respiration Physiology 109 (1997) 177–194The elephant’s respiratory system: adaptations to gravitational stress R.E. Brown a , c , J.P.

Respiration Physiology 109 (1997) 177–194 The elephant’s respiratory system: adaptations to gravitational stress R.E.

The elephant’s 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, Har ard Uni ersity, Boston, MA 02115, USA b Department of Anesthesia, DA 717, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA c Zoological Institute, Department of Zoomorphology, Go¨teborg Uni ersity, Medicinaregatan 18, S-413 90 Go¨teborg, Sweden

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 elephant’s respiratory system and speculate on their functional significance. A distensible network of collagen fibers fills 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 fluid that may replace the gravity-dependent flows of smaller mammals. The lung is partitioned into 1 cm 3 parenchymal units by a system of thick, elastic septa that ramify throughout the lung from origins on the lung’s 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 influ- ence 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

influenced 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 con- cerning 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 S0034-5687(97)00038-8

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1913; Earles, 1929; Bu¨ßgen, 1988). Descriptions of attachments between lung and chest wall in ele- phants (connections form in late fetal life, Earles, 1929) have been in the literature since 1682 (Moulin (1682) cited by Earles, 1929). Early spec- ulations 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 elephant’s need for forceful inspira- tory 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-di- aphragmatic 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 be- tween 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 elephant’s respiratory system. We suggest that the morphology of the elephant’s lung indicates that gravitational forces do indeed influence 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 breath- ing. Rather, the loose pleural space connective tissue may be important in the regulation of the flow of pleural fluid. In the elephant the delicate gas exchanging pulmonary parenchyma is compartmentalized into grape-like units, 1 cm 3 , by an extensive system of thick, elastic septa that originate from the lung’s external capsule and which ramify throughout the lung. We suggest that the par- enchymal compartments are suspended by the elastic septa so that dependent areas of the lungs are not compressed by the lung above, and non- dependent alveoli are protected from overexpan- sion, thus reducing the effects of gravity acting on the elephant’s 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 find- ings were encountered during the necropsy begun immediately after death.

2.2. Thoracic dissection and gross obser ations

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 di- aphragm’s 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 first to last rib, sternum to the dorsal costal arch. Assessment of the mobility and deformability of the pleural connective tis- sue 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 superficial, of both lungs. A primary bronchus and one of its branches was serially sampled from where it en- tered the lung’s hilum until it reached a diameter of 1.0 cm. A section of mid-trachea (intratho- racic) with five complete rings was preserved in glutaraldehyde.

2.3. Tissue preparation

2.3.1. Fixation Tissue specimens were rinsed with 0.05 M phos- phate buffer (pH 7.2) and fixed 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 fixative was replaced twice within the initial 4 h. Some tissue blocks were stored in glutaraldehyde for later sub-gross dissec- tion. Tissues for electron microscopy were post- fixed in 1% osmium tetroxide in 0.05 M cacodylate buffer (pH 7.2). All tissues for micro- scopic examination were dehydrated in a graded ethanolic series (70% to absolute). The elephant’s 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.

at necropsy and immersed in 10% buffered neutral formalin. Fig. 1. Diagram of a section through

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 filling 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.

2.3.2. Light microscopy

Specimens were processed according to stan- dard paraffin techniques. Sections, 6 m thick, stained with hematoxylin and eosin, Verhoeff-Van Gieson’s and Masson’s Trichrome were pho- tographed with a Nikon photomicroscope. Montage reconstructions were accomplished using overlapping fields digitally photographed (10× ) with a Dage 725 CCD camera on a Leitz photo- microscope into a Zeiss IBAS image analysis sys- tem. Individual fields were reassembled using Adobe-Photoshop ® running on an Apple Macin- tosh, Quadra 605.

2.3.3. Transmission electron microscopy

Specimens were embedded in Spurr’s resin ac- cording to standard procedures. Thin sections, 100 nm, stained with uranyl acetate and lead citrate were examined with a Phillips 300 electron microscope.

3. Results

3.1. Gross obser ations

3.1.1. The animal

The maximum external dimensions of the tho- rax (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) sup- ported by one knee (6–8 cm 2 of contact) on the distal end of the trunk did not collapse the en- closed nasal tubes upon the researcher’s fingers inserted into the external nares.

3.1.2. Thoracic ca ity

A small (15 cm) initial incision through the diaphragm’s central tendon immediately resulted in a large pneumothorax (Fig. 1). Surrounding the pneumothorax cavity was a lustrous, smooth sur- face 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

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caudo-dorsal rib cage (Fig. 1B), the PSCT sur- rounded 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 filled all the space between chest wall, fully collapsed lungs, di- aphragm 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 finger. Handling and dissecting the unstressed PSCT was difficult, due to its slickness and the ease with which its ‘substance’ was sheared when attempting to pene- trate it. Examination of the lustrous PSCT at- tached to excised lungs and chest wall revealed it to contain myriad small spaces (fluid and air filled) among the wet, shiny fibers; the gross ap- pearance 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.

3.1.3. Diaphragm The diaphragm’s insertion extended from the ventral end of the first pair of ribs caudo-dorsally to the most dorsal aspect of the 21 st (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 connecti e 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

from being elevated perpendicular to the inner surface of Fig. 2. Diagram of a 25 ×

Fig. 2. Diagram of a 25 × 25 cm slab of the elephant di- aphragm’s central tendon lying on a surface with the attached pleural space connective tissue uppermost. The surperficial 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.

the rib cage. With the diaphragm held perpen- dicular to the rib cage, the attached PSCT was not taut, but remained loose and easily de- formed. 2. A 3 × 3 cm section of the central edge of the diaphragm muscle was isolated from sur- rounding 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 di- aphragm’s insertion on the rib cage, over a distance of 26 cm. 3. A slab of the diaphragm’s central tendon, 25× 25 cm, was isolated without displacing the attached PSCT and placed with its abdom- inal side down (Fig. 2). At the midpoint along one edge of the slab, the most superficial (clos- est to lung in vivo) portion of the PSCT was grasped with forceps. The point of the forceps grasping the PSCT could be easily moved to- wards the opposite side of the slab, creating a deep ‘V’ shaped pattern in the superficial 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 superficial layer was released it remained in its distorted ‘V’ configuration, retracting towards its original position by less than 2 cm.

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et al . / Respiration Physiology 109 (1997) 177–194 181 Fig. 3. Elephant lung, superficial relationships

Fig. 3. Elephant lung, superficial relationships of macroscopic connective tissue components, light microscopy reconstruction, lung fixed at zero stress. The lung’s 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 lung’s external capsule. The distensible intralung septa (ILS) originate from the lung’s external capsule and ramify throughout the pulmonary parenchyma, dividing the parenchyma into compartments 1 cm 3 .

After

removing

the

lungs from the thorax,

length and when released returned to their origi-

PSCT attached to the lung could be easily stretched to considerable lengths. A handful of

nal highly folded configuration (Fig. 3). From the external capsule strong, elastic connective tissue

PSCT lying next to the lung in the unstressed state could be easily stretched uniaxially to 20–50

septa ramified throughout the pulmonary par- enchyma. All grossly visible intrapulmonary air-

cm

without disturbing (elevating) the surface of

ways were invested with a thick coat of dense

the

lung itself. The elongated ‘stem’ of PSCT

connective tissue. The airways with their peri-

raised from the lung surface, when released, did

bronchial connective tissue gave the cut surface of

not

reform itself into the smooth layer covering

the lung a very rough, pebbly feel and appeared

the

lung, but instead remained in a deformed,

as if the walls of all airways contained abundant

elongated blob lying on the surface. The PSCT

cartilage, although no cartilage was found on

was a very slimy material that stuck to most

subsequent microscopic examination. When the

things it touched (e.g. latex gloves, table tops) but

cut ends of small, 2.0 mm luminal diameter

was

difficult to grasp because it was so easily

airways were grasped with forceps considerable

sheared (deformed). A layer with an unstressed (but aerated) thickness of 5 cm could be easily squeezed between thumb and finger to a layer 1

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

mm

thick.

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 at- tached to this capsule, which is homologous with

the visceral pleura (albeit without a serosa, see

below) covering the lungs of other animals. Iso- lated strips of the lung’s 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 ar- rangement of the tracheal rings, the overlapped, alternating attachments making a saw-tooth pat- tern (zipper like) along the trachea’s dorsal sur- face. The dorsal third (both sides) of each ring

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et al . / Respiration Physiology 109 (1997) 177–194 Fig. 4. Elephant lung 15 cm deep

Fig. 4. Elephant lung 15 cm deep from its pleural surface showing macroscopic relationships of connective tissue components, light microscopy reconstruction, fixed under zero stress. The distensible intralung septa (ILS) ramify extensively throughout the lung, dividing the pulmonary parenchyma into compartments 1 cm 3 . 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 0.1 mm are attached within and surrounded by the supporting framework of the intralung septa. When this figure 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 lung’s 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 flattened oval cross section laterally, ending with irregular facets for their double articulation dorsally. The rings were fixed to one another in the overlapping areas and across their articulations by tough fibrous tissue. The tracheal mucosa had many 1 mm deep longitudinal rugae. These rugae could conceivably result from constriction of the trachea by contrac- tion 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 fibrous tissue fixing 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, fixed in glu- taraldehyde were dissected and microscopically examined at 10–40 × . The fixed PSCT, though less distensible than when fresh, could be easily deformed and elongated to more than twice its fixed-resting length. The PSCT appeared as a complex, 3-D fibrous network containing myriad air- and fluid-filled spaces 0.1 mm 3 ; the spaces between fibers occupied a larger volume than the formed elements. In most areas the collapsed lung’s external cap- sule was finely folded or pleated (Fig. 3). The connective tissue septa seen on gross dissection partitioned the pulmonary parenchyma into iso- lated compartments of about 1.0 cm 3 (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 fixed parenchyma could be easily scraped from the septa leaving strong- walled 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 ves- sels 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 (lo- cated within an adjacent intralung septum) and

vessel (lo- cated within an adjacent intralung septum) and Fig. 5. Segement of elephant trachea taken

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 trachea’s lateral aspect (see cut off rings, cross-hatched in (A). The overlap is reinforced with a dense fibrous 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 fibrous aponeuro- sis. With a saw- tooth-like appearance, the end of each carti- laginous 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 fibrous tissue (FA in C). Note that the cross-sectional shape of the cartilagenous rings changes con- siderably as one moves around their circumference. The tra- cheal mucosa (M) is strongly pleated into numerous longitudinal rugae.

coursed to the center of the parenchymal com- partment before giving off a cluster of terminal branches. These compartmental branches and their terminal ramifications (to the limit of micro- scopic observation) were covered by a reflection 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 lung’s external capsule deep to the PSCT, the PSCT and the internal chest wall and diaphragm (Fig. 6A and 6C). How- ever, we found a normal appearing serosa invest- ing the abdominal surface of the diaphragm. The PSCT, including its attachments to the external lung capsule, diaphragm and internal sur- faces of chest wall, was almost entirely collage- nous with but an occasional elastin fiber ( 1 per 40× field) 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 lung’s external capsule. Whereas in most of the PSCT there was no apparent organi- zation of the collagen fibers of the PSCT (Fig. 6A and 6C), in some areas small bundles of collagen fibers 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 negligi- ble metabolic needs of this collagenous tissue. Dispite the much greater density of formed ele- ments in the lung capsule relative to the pleural space, there were about 7-times as many vessels per field in the loose PSCT, 7.5 per field, 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 tis- sues of the diaphragm’s central tendon and costal periosteum showed 0.3 vessel per field.

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et al . / Respiration Physiology 109 (1997) 177–194 Fig. 6. Elephant lung, light microscopy, Verhoeff-Van

Fig. 6. Elephant lung, light microscopy, Verhoeff-Van Gieson’s elastin stain, collagen red, elastin black, smooth muscle brown. (A) Filling the center of the figure is the dense and highly distensible external capsule of the elephant’s lung. The external capsule has an elastin (black) content of at least 35% relative to that of collagen (red). Across the top of the figure, external to the lung’s 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 elephant’s lung or within the pleural space connective tissue. The pulmonary parenchyma along the figure’s lower edge is divided by one intralung septum originating from the lung’s external capsule. Compare the thickness of the elephant lung’s external capsule in this figure 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 (identified 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 fibrous tissue. The lung’s external capsule crosses the lower, left corner of this figure. (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 com- posed of 35–50% elastin and 50–65% collagen (surface area approximations) (Figs. 3 and 6AFig.

7). Intralung septa, with a fibrous arrangement similar to that of the lung’s external capsule, were continuous with the fibrous 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 lung’s 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 field as did the domestic animals examined for this study, al- though 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 fibrous tissue of a septum was continuous with the fibrous tissue forming the wall of the vessel or airway. The fibrous tissue layer sur- rounding airways and vessels contained copious amounts of elastin external to the muscularis (Fig. 8A). Larger vessels and airways were often lo- cated within the junction of two or more septa.

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

cartilaginous plates could be identified, the intra- Fig. 7. Elephant intralung septa, transmission electron mi-

Fig. 7. Elephant intralung septa, transmission electron mi- crograph. The highly distensible intralung septa are composed of 35% ( 50% in this view) amorphous elastin fibers (E) with the remainder being bundles of collagen fibers (C) and the occasional fibroblast (upper left). Bar =1 m.

pulmonary airways were devoid of cartilaginous tissue. The latter was surprising considering the pebbly feel of the lung’s cut surface and the fact that cartilage is a prominent intrapulmonary air- way component in other species, including the horse, cow and pig. A circumferential sheath of dense connective tissue, continuous with and identical in appear- ance 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 airway’s septal sheath from its thick wall. Septa radiated from the sheath in the plane of the airway’s axis and continued into the pulmonary parenchyma. Smaller airways ( 2 mm luminal dia) were always tightly invested within a septum (Figs. 3, 4 and 8A). The fibrous tissues of the airway and associated septum combined to create thick air- way walls whose thickness external to the mucosa was 30–200% 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, ax- ially 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 sufficient 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 film of amorphous material, assumed to be mucus, coated the mucosa in most airways and rarely polymorphonuclear cell were found ( 1 per 40 × field). The submucosal and muscu- laris layers of the airway walls contained an abun- dance of elastin fibers relative to that of the other taxa examined (Fig. 8A and 8BFig. 9). The orien- tation of the submucosal elastin fibers was pre- dominantly axial (mostly cross-sections of fibers were found in airway cross-sections), with the elastin fibers of the submuscularis and attached intralung septa oriented more nearly radially (lon- gitudinal sections of fibers found in cross-sections

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et al . / Respiration Physiology 109 (1997) 177–194 Fig. 8. Elephant lung, light microscopy, Verhoeff-Van

Fig. 8. Elephant lung, light microscopy, Verhoeff-Van Gieson’s elastin stain. (A) Smaller conducting bronchus with attached septum. The mucosa of the airways in these lungs fixed at zero stress was always found highly pleated. The majority of the submucosal elastin fibers (black) have an axial orientation, in contrast to the elastin fibers 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 fibers lying deep to the airway’s mucosa (arrowhead) continue into the alveolar duct (top 3/4 of figure). The pulmonary parenchyma is highly distorted and collapsed in these lungs fixed at zero stress (relaxed). Note the magnification of this figure is twice that of all other color micrographs. (C) These three micrographs represent one complete and continuous slice through the diaphragm’s central tendon from abdominal (1) to thoracic (4) sides (about 10% of the central tendon’s 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 fibers (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 fibers. 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 elephant’s much thicker and denser external lung capsule seen in Fig. 6A. In addition, the elephant’s external capsule contained much more elastin. All micrographs of external lung capsule, e.g. Fig. 6A and D of this figure were stained simultaneously and are shown at identical final magnification.

of airways). The submucosa of terminal bronchi contained abundant, axially-oriented elastin fibers 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 fixation; thus the fine alveolar architecture was distorted or obscured

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et al . / Respiration Physiology 109 (1997) 177–194 187 Fig. 9. Intralung septa tethering a

Fig. 9. Intralung septa tethering a large airway. All airways had one or more intralung septa (ILS) attached to, and radiating normal to the airway’s 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 fibers and adipose tissue (peribronchial tissue). The submucosal layer contained large numbers of predominantly axially oriented elastin fibers, seen here as a thick dense-black band deep to the airway’s pleated mucosa (M).

(Fig. 6DFig. 8BFig. 10). Many collagen fibers and bundles of collagen fibers were found immediately beneath the pulmonary epithelium and deeper within the alveolar membranes (Fig. 10). The apparently increased collagen content notwith- standing, the elephant’s alveolar membranes, type 1 and 2 pulmonary epithelial cells and capillaries, appeared like those of other mammals. Gas ex- change tissue lined the intralung septa.

3.3.5. Inflammatory cells

Alveolar macrophages were found within the elephant’s lung. Mononuclear cells found within alveolar capillaries have provisionally been iden- tified as pulmonary intravascular macrophages (Fig. 10).

3.3.6. Diaphragm

The central tendon over 3 mm thick was com- posed of three distinct layers of dense, collage-

nous tissue. The layer nearest the abdomen also

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

3.4. Comparati e morphology

In conjunction with the present study the lungs of domestic cow, horse, and pig were examined histologically (Fig. 8D). In these domestic ani- mals, the lung’s external fibrous capsule was al- ways less than 0.18 mm thick (elephant’s capsule was 0.25–1.75mm thick) and only rarely were elastin fibers identified (elephant’s capsule was elastin rich). The intralung septa of these animals were commonly composed of two thin collage- nous sheets, which together were always less than 0.08 mm thick (not counting any gap between layers), and only very rarely contained elastin

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fibers. In contrast, the elephant’s elastin rich in- tralung septa were found in a range of thicknesses up to that of its external lung capsule. The inter- lung septa of the domestic taxa were only infre- quently found to attach to an airway. In sharp contrast to those of the elephant, the larger intra- pulmonary airways of these domestic animals contained cartilaginous rings while smaller, non- respiratory airways contained cartilaginous plates. The thickness of the walls of smaller airways in these domestic animals was 25% of the airway’s submucosal diameter (elephant’s airway walls were 30–200% of submucosal diameter). Relative to those of the elephant, the wall of the airways of these domestic animals contained very few elastin fibers.

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 addi- tion, except for examining the mobility of the PSCT, we have no in vivo or in situ experimental measurements of the mechanics of the elephant’s

measurements of the mechanics of the elephant’s Fig. 10. Elephant’s alveolar septum, transmission electron

Fig. 10. Elephant’s alveolar septum, transmission electron micrograph. Note the large quantity of collagen fibers and bundles of collagen fibers (arrowheads) lying between the pulmonary epithelium (PE) and capillary endothelium (CE). We have provisionally identified the large mononuclear cell filling the capillary lumen as a pulmonary intravascular macrophage. Bar =1 m.

respiratory system. The histologic findings re- sulted 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 connecti e tissue

4.1.1. Lung mo ement relati e to chest wall during breathing The mobility, extreme compliance, ease of shearing and slipperiness of the pleural space con- nective tissue (PSCT) implies that it has negligible influence on the sliding motions of the lung rela- tive 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 inflated configuration 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 elephant’s 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 di- aphragm, functioning identically to the liquid- lined pleural space of other mammals. Unique to the elephant’s pleural space is that it is finely divided by the fibrous continuum of PSCT, the fibers of which are lubricated by the elephant’s pleural fluid. We consider the deformability or stretchiness of the PSCT to be a function of the network organi- zation of its fibers which are almost entirely in- elastic collagen. While the ease with which it can

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be sheared is, quite frankly, amazing, there are other examples (different taxa and different anatomical locations) in which networks of colla- gen fibers undergo large physiologic deforma- tions. A common example is the subcutaneous tissue attaching skin to underlying muscle fascia, e.g. the domestic cat’s thigh and knee can move cranio-caudally 15 cm beneath the skin of the flank without disturbing the overlying skin.

4.1.2. Ele ating water ia the trunk during drinking Speculations in the literature suggest that the connections between the elephant’s lung and chest wall, i.e. PSCT are necessary to prevent a sponta- neous pneumothorax and pleural vessel hemor- rhage from occurring secondary to either elevating water in the trunk for drinking or force- ful inhalations against a high resistance to flow through the long trunk (we consider a high resis- tance to airflow doubtful but no measurements are available) (Todd, 1913; Earles, 1929; Short, 1962; von Beyer et al., 1990). Large trans-respira- tory pressure differences (from alveolar space to body surface), up to 300 cmH 2 0 in a large ele- phant completely filling 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 fill 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 pul- monary 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 elephant’s transpulmonary pressures are unaf- fected by drinking and thus the likelihood of a spontaneous pneumothorax (breech of pleural in- tegrity) is not only no higher than that of other animals but probably much lower because of the elephant lung’s very thick external capsule. Fur- ther, cavitation (i.e. separation of tissues due to extremely low pressure) within the elephant’s pleural space secondary to large transthoracic pressure differences cannot occur at absolute pres- sures above 50 torr (710 torr relative to atmo-

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

4.2. Mechanical implications of ele ating water in a long trunk

However the extraordinarily low intrapul- monary air pressures, — 300 cmH 2 O, required to elevate water to a height of 3 m in the nasal tubes with respiratory muscles could potentially (1) col- lapse 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 possi- ble only if the extra-thoracic trachea and nasal tubes within the flexible 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 ossified tissue are a function of the trunk’s thickness. An indication that the trunk is sufficiently resistant to collapse is that a local compressive stress 6.0 × 10 3 cmH 2 O failed to collapse the distal nasal tubes where the trunk- wall 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 be- tween adjacent rings. The intrathoracic trachea, like the lung, is not affected by the trans-respira- tory 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 modifications within the elephant’s 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 elephant’s diaphragm that may assist in the production and support of those forces. The thick, well developed, trilaminar structure of the elephant diaphragm’s central tendon appears to

be an adaptation for the support of large trans-di-

aphragmatic pressures (Fig. 8C). Sacks and Chuong (1992) describe the organization of the

collagen fibers of the single layered central tendon

of other mammals as orthotropic (having parallel

fibers; see also Griffiths et al., 1992). There is a

weakness with such an arrangement in that a

rupture or separation could occur between fibers

at forces far below those required to break the

fibers themselves. Whereas the collagen fibers of each individual layer of the elephant’s central tendon are also in a parallel arrangement, the different alignments of the fibers in the three

layers of the elephant’s central tendon would con- fer nearly isotropic mechanical properties to this trilaminate structure. The elephant’s 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 ventrodor- sal= 143 cm) we can calculate the oval circumfer- ence (388 cm) and axially projected area of the diaphragm (Area di ) to be 1.12× 10 4 cm 2 . 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 (CSA M ) to be 1.16 ×

10 3 cm 2 . (This underestimates both projected area and muscle cross-sectional area because of the oblique orientation of the elephant’s diaphragm.) Assuming the elephant’s diaphragmatic muscle generates a maximal stress ( ) (tension) similar to other animals’ skeletal muscles (2×10 6 dynes/

cm 2 ; Powell et al., 1984), we calculate the maximal

pressure (P di ) that this elephant’s diaphragm can produce is

P di = M × CSA M Area di

The orientation of the surface of the elephant’s 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 in- volved in drinking, or an adaptation for the sup- port of the static pressure gradients associated with gravity acting on the very different densities of thoracic and abdominal viscera. These possibil- ities remain uncertain and deserve further study.

200 cmH 2 O

4.3. The elephants respiratory system:

adaptations to gra itational stress

The question, ‘Do gravitational forces set limits on the design of the respiratory system?’ (Leith, 1976), has been long standing in respiratory phys- iology. At least two features of the elephant’s respiratory system appear to be structural adapta- tions 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 in- creases 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 inflation existing at low lung volumes resulting from the compressive effects of gravity, but that as lung volume is increased the regional differ- ences would disappear due to a ‘stiffening effect’ due to the parenchyma’s 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 inflation to occur in the more dependent and less distended regions of the lung.

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In the elephant, the pressures inflating the alve- oli would vary by 50 cmH 2 O 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 = 0.35 gm/cm 3 ; see Ganesan et al., 1995). As- suming the elephant’s lung has a pressure-volume relationship similar to that found in other mam- mals in which the vital capacity spans a transpul- monary pressure range of 25–30 cmH 2 O, the 50 cmH 2 O 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 physio- logically normal distending pressures during breathing. It follows that at all physiologic lung volumes, the elephant’s tall lung, without struc- tural modifications, would have a large propor- tion 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, P pl , which supports the whole lung in gravity, and the verti- cal gradient in distending pressure of the pul- monary parenchyma, which we argue for the

elephant must be substantially less than P pl . In the lungs of other (smaller) mammals it is argued that the pressure distending the parenchyma is the

elastic recoil pressure of the lung, P el

(Mead et al., 1970) where P A is alveolar pressure. This pressure is also the stress within the par- enchyma averaged over an area transecting many alveoli (e.g. several mm 2 ). In the elephant this assumption may not hold; within the small par- enchymal units stresses averaged over several mm 2

may be substantially less than the stresses within the lung and its supporting intra-lung septal sys- tem averaged over many cm 2 . At least in very large lungs it appears unreasonable to expect lung tissue to be self supporting. If the mass of par- enchyma, blood supply and airways comprising the elephant’s lung were to be supported solely from fine internal structures at the alveolar and ductal level it seems reasonable to expect consid- erable reinforcement within the lung. In fact, ele- ments capable of supporting compressive stress would be necessary within its dependent portions. Such a structural adaptation at the level of the

=P A P pl

( L )

alveolar membrane has not been found, or not recognized, in large mammals (including the ele- phant). We suggest that the elephant lung’s capsule and intralung septal system is a structural mechanism that supports parenchymal, vascular and airway structures so that the alveoli are functionally iso- lated (partially or totally) from the gravitation effects described above. The thick external capsule (visceral pleura) of the elephant’s 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 ramification of the intralung septa throughout the pulmonary parenchyma serves as a space filling, interconnected set of surfaces tessellating the lung and supporting the resulting small ( 1 cm 3 ) me- chanically isolated compartments of parenchyma. The high elastin content and coextensive net- work of collagen and elastin fibers found in the elephant lung’s intralung septa (and external cap- sule) (Fig. 6A and 6DFig. 7) is very similar to that found in mammalian Lig. nuchae and the bird’s 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 com- posite forming the structural elements of the ele- phant lung’s external capsule and intralung septal system would allow these structures to maintain nearly constant support of the pulmonary par- enchyma 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 expan- sion. That is to say, if pre-stress of the septal system at low lung volume were such as to sus- pend parenchymal compartments uniformly, there would be little regional difference in the inflation of the elephant’s alveoli over a wide range of lung volumes. Indication that prestress conditions do exist within the elephant lung’s highly elastic ex-

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ternal capsule and intralung septa is reflected in their tightly folded (pleated) conformation seen in the deflated, relatively gas free lung and in iso- lated tissue blocks. The linear elastic support mechanism provided over the entire lung by the elephant lung’s 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 elephant’s pul- monary parenchyma within a single parenchymal compartment to be similar to that of other mam- mals 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 elephant’s vital capacity could span a range of 25–30 cmH 2 O as found in other mam- mals. However, if the elephant is to experience ventilation of all its alveoli, its lung-wide mean distending pressure (as well as P pl ) must be higher than other animals, at least 50 cmH 2 O. Over a 7-fold range of body mass there ap- peared 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 al- most 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 elephant’s 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-support- ing in the fashion described by Milic-Emili et al. (1966) and West and Matthews (1972). This tran- sitional size appears to be one at which the differ- ence in average recoil pressures between the top and bottom of the lung approaches the difference in recoil pressures between relatively un- inflated and fully-inflated parenchyma. The very sparse ( 5%) elastin content of the intralung septa of the pig, cow and horse (and many other mam- mals, data not presented here), relative to that of the elephant (35–50%), 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 alve- olar septa at high stresses.

4.3.2. Elastin content in walls of large airways and essels The walls and investing septa surrounding the elephant’s intrapulmonary airways and vessels contain many axially oriented elastin fibers (Fig. 8A). If the extensibility of the elephant’s intrapul- monary 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 intrapul- monary mechanisms exist to isolate and protect the bronchial tree from axial distortion, we would suggest that the elephant’s elastic bronchial tree undergoes considerable length changes. Lai-Fook (1979) indicates that the larger arteries within the lung, function to limit regional expansion of pul- monary parenchyma, contributing to the pul- monary interdependence described by Mead (1961) and Mead et al. (1970). However, the dense elastic coat of the elephant’s pulmonary vascula- ture and the septal tissues investing it indicate that these vessels are quite extensible and that they probably lengthen as the surrounding tissue ex- pands, contributing to local pressure-volume characteristics without limiting expansion.

4.3.3. Flow of pleural fluid Pleural fluid, under gravitational influences, ex- hibits 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; Miseroc- chi et al., 1988). Departures of liquid pressure gradients from 1 cmH 2 O/cm vertical height have been associated with viscous pressure losses aris- ing from pleural fluid drainage (Lai-Fook and Rodarte, 1991) and may also be involved with ‘pumping’ of pleural fluid secondary to oscillating shear stresses during breathing (Butler et al., 1995). We suggest that the fibrous matrix of the

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PSCT (Fig. 6A, 6B and 6C) may be necessary to retard gravitationally driven flow of pleural fluid from top to bottom of the elephant’s tall lung, which could result in excessive thinning of the lubricating layer of liquid between lung and chest wall and collections of pleural fluid in the ventral thorax. The myriad small spaces within the fibrous continuum of the PSCT may act to slow the flow of pleural fluid via a process of percolation through the fibrous matrix. Thus, the pleural fluid would act as a lubricant between lung and chest wall by lubricating the motion between adjacent fibers of the PSCT. The abundance of small ves- sels and lymphatics (Fig. 6B and 6C) throughout the PSCT suggest that large local fluxes of pleural fluid occur within the tissue’s fibrous matrix. Thus, in contrast to the pattern of pleural fluid flow in other mammals with a fluid filled pleural space, the elephant’s pleural fluid may be pro- duced and reabsorbed locally, reducing or elimi- nating the flux of fluid from top to bottom of the thorax.

4.4. Maintenance of airway patency and the elephants intralung septa

During expiration, when the pressure driving flow is coupled to peribronchial pressure, flow limitation could potentially occur in any unsup- ported intra-thoracic airway. In contrast to other mammals, the elephant has no cartilaginous sup- port of its intrapulmonary airways to maintain airway patency. However, the elephant’s intralung septa which surround and attach to airways larger than about 2 mm luminal dia. radiate normal to the airway’s axis and ultimately connect to the lung’s external capsule, suggesting that the lumi- nal patency of these larger airways may be main- tained 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 tether- ing 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 relation- ship 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 bronchi- oles airway patency must be maintained solely by the local radial tethering forces of attached alve- olar membranes in a fashion identical to that of other mammals (Fig. 8B). Many small bronchi, 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 pro- duce 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 rela- tively gas-free state of the intact lungs following their removal from the thorax was evidence that small airways remained patent at very low dis- tending pressures. Interestingly, the folding pattern of the ele- phant’s 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 broncho- constriction produced by activation of airway smooth muscle. With reductions in airway caliber to the degree responsible for the observed mu- cosal folding, there would be substantial increases in airway wall thickness. It is possible that the elephant’s 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 com- pression of the elephant’s 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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demands for parenchymal support, but that with attachment to the airways, it subsequently re- placed the need for cartilaginous support of air- ways’ lumens. The presence of cartilaginous rings supporting the lumens of the elephant’s extra-pul- monary bronchi and trachea demonstrates that elephants are indeed capable of developing such airway support structures.

Acknowledgements

We thank Professor Kurt Benirschke for intro- ducing us to this interesting problem, for his encouragement and for discussion of the ideas presented here. Discussions with Drs Fred Hop- pin 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 figures. 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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