The Natural History of Atherosclerosis

By MARCEL M. VASTESAEGER, M.D.,

From the Centre d 'hude des Maladies des Arteres

Coronaires, Department of Cardiology and Department of Clinical Pathology, Hopital d 'Ixelles,
Brussels, Belgium.
1962

R. DELCOURT, M.D.

term when alluding exclusively to one or another of the various lesions of this condition.
Such habits have made literature more and
more confusing.
As a matter of fact, the only valuable way
of defining each kind of arterial degenerative
lesion is based on histologic criteria. This is
why, referring to etymology, and according
to the recent nomenclature of the World
Health Organization,3 we shall confine ourselves in this paper to the following definitions:
1. Intimal sclerosis: diffuse fibrous thickening of the intima, generally associated with
degeneration of the intimal elastic membrane
and with sclerosis of the mnedia. Grossly, diffuse intimal sclerosis gives rise to some dilatation and to lengthening (unrolling) of the
involved vessels, but the lumen is not impaired.
2. Atheromna: strictly focal lesion (e'tVa' =
tumor ), characterized by the presence of
lipids (a'Thj'py = mush), conferring to the
plaque a grossly yellowish color.
3. Atherosclerosis: more recent word introduced by Marchand4 and referring to a lesion
intermediate between intimal selerosis and
atheroma. Usua.lly focal, atherosclerosis has
an important fibrous component and constantly shows lipid deposits. In their advanced stages, atheroma and atherosclerosis
grossly impair the lumen of the vessel.
As a rule, atherosclerosis is less localized
in animals than in man. Especially in birds,
stenosing atheroselerosis is much more diffuse
than usually observed in human arteries
(fig. 1).
The diffuseness of spontaneous atherosclerosis in animals often makes it very difficult
to establish a clear-cut difference between
mere intimal sclerosis and the first stages of
atherosclerosis. This explains why it is very

A LTHOUGH unceasing attempts have been

j L made to increase the knowledge of human and experimental atherosclerosis, the
pathogenesis of this disease is still far from
being thoroughly understood. The uncertainty
of present conceptions concerning atherogenesis is emphasized by the rapid succession
during the last 20 years of several theories
allotting a predominant role to different substances such as cholesterol, animal fats, saturated fatty acids, and nonessential fatty
acids, and the series has probably not yet
come to an end.
Hitherto, epidemiologic investigations have
been quite anthropocentric: although the incidence of clinical atheroselerosis among various human populations of the world is relatively well known, the frequency if its
spontaneous occurrence in the entire group of
animals having a specialized circulatory system is almost completely unknown. In other
words, most of the natural history of atherosclerosis is still to be discovered. The purpose
of this paper is to sum up the fragmentary
knowledge in this field, and to pose some
problems the solution of which might be of
interest for the etiology of atherosclerosis.
In order to avoid confusion, it may be opportune to define once more arteriosclerosis
and atherosclerosis. As, proposed by Lobstein'
arteriosclerosis is a generic term, designating
several arterial degenerative processes giving
rise not only to a thickening, but also to a
loss of elasticity of the vessel wall (diffuse
intimal sclerosis, Mdnckeberg 's2 medial sclerosis, focal atheroselerosis, etc.). Unfortunately, too many workers have discarded the
generic sense of arteriosclerosis and used this

Circulation, Volume XXVI, November

AND

841

V4 YASTE,SALENE,1)ELCU(R)IlZ

1;42)

thIeI filst to shio 1ht.v

eI.Xot iaInIIIa1IIIils a 11(I
ibirds livilig i i cat it v t i viii c1ila d I t,
(legelereati_V(e arte-rial li1siols. (ouit --11111g'. th10
wock of' Fox, Baltt(iffe
1
( 0rmihi
)
showed
that tle ineideliee (,f (legCel,ieatvx' aterial
na
phleC tllo

seevs; toio

e(aS( lrO-flnrt jioallxt.v

xv.itl tile rate of' over 1l)opllation of zoos, and(i

Aw-itlh the str'esses t-o wheli tI-le animials arie

Su1)4j et(ed. Eeei itlY, Ealct -lifit'c'-,Yerasi II idc1(7

ani-d lE'lliot t' iinsisted upIol I lie, icnireasi ig Iecuei(eyll(v of steioEsiIg eorounrx lesimns Hi tlie

Ihomel(othernic anIimal's of the Philadelhlia

1950, inlvoking agai(n aIs Iet
l Cause
[avorllou the vxol iti loll of t hese elsioiis I lie
'V lug(Ieist o ot
tne ej
ivet animal] plop(,t]phv

ZV0O since

lat.ioll.

In1- the oi-ller hanilld , spoiiau leons Itt inal

selerosis or athlcrosrlIteosis lhas beent. dese Ins
in iny species of domnestie ail;fmoals, 1lir(ll aild
ni.iiiiiials, alike i(ihiekeli
ill)it,1 d{log,11' 12
and pigeoli1.1" So,on. einax oiisiilde it fils1111v
established thfiat mI(ost sp)cies o fiomeotl)hrlni'
animals that: ni has doinest:hafite.d orl reduced
to Car)tivity, Jim
ov s)oiltxtleousl.x (lexeh)l) 50m111
towin of arCCi 1)8(1
serss

Coroner!

artery

of

javaniicus

Figure 1
(otl
jeeon tee

duck

(Dendro

Ahor.]). Coneentrie thickmug of the itirln : the

1(9 1 stiocytes wuithi
hinun
riletr cytoplasm sh/io the atheroselerotie natun'C

cygnus

ofthi's lesion.
the same

ness

Gou

A.

X
>2t7. B. Another section. in

erterilq shoo-ingfJ importance

of the lipid deposits. Stdorla

.\Vith]54,ia

fhil)-potauitt l).oiiits ieeessary forL the coiupllet'e

11iderstaialidl1 (of thle re.nesis of altelial de
(rei('Lerativte
ee esses
es a
mid of
eni ntIatull
listolry TeMaiJi Ob'senrilet.
tuinde:r thie mIpi(s s of tilew il,iegiaj (celer
fror theC Studly ()f (`oronary- Artery I)iseasc i
svstemiatic researelh jilto) the arter ial degeeIra.eltire ma-ifestat iolXs ii the anlinial. killn-dom
hans b:ceen uniIdertakecn for a fe xvv e ari-s. I I viewv

tidt (liffntst-

bleeak B X27

difficult, if niot im-possitAble, to dissociate the

natur al history of atherosclerosis from the
nataural history of dliffuise intiinial selerosis.
The Avor k carried out for niearlyA half
cen.jturv at the Zoo of Philadelphia has built
mtiost of our present klnowledge of spontaneouLs
atheroselerosis in wil(d aniimals. Fox,5'6 was
a

*Tlese stuSdies couu(II T C8

( cir
1 I idout only bec.lilso
of the Very obliging hell) of 1m1aty11y oigaiizit-.tioalls iln
differenit coiiitries. Aee wish to thank sotouig otlcrs:
The Intrn
1a
telt;iolall Soce; Wtv of (ald-iolog) (I)r-s. White
no]l Kat.tz); Tf e Aiitwerp) Zoo (Mr. V.an den Bergl
aInld Pi-of. Morhimatl-ins); Thie ''l Justiituluto Latinio
Aincrica:o de \Arttomia patologi,jc II of Me'eico (PIr of
Costero); The Laboratory of Paithology of the
lrnw;sels 'University (Prof. Distiai aniid Dr. Pavine ni
tier); The med(ical Laboratory of Stanfle) v lle (CoII-

go); Thie capturing camp of Einpulti (Conlgo), Mr.

d(o AMediiia; The Commilission of Coopa-trative Alnltomy
(Kaiisas, USA), Dr. A[aizl<e; Tue Scienitific Peseairebl
ilnstitulte of Pilill,itz (Fiancec), MAr. Bari6ty and
AMr. PIcrcier; The MArlrine Biology Stationl of l?oscotfi
(France) ; Thle ArAsterdani Zoo Toil Veteriina.ry School
of Utrieclit s Uniiversit3, Prof. Zw art.
Circulation, Volumc XX V., Novmnber 1.962

NATURAL HISTORY OF ATHEROSCLEROSIS

of the huge field to be explored, these investig-ations are still in their first stages, and the
specimens hitherto examnined of invertebrates
are too few to fuirnish valuable data. Among
vertebrates, however, a few points worthv of
initerest alreadv have been recorded.

Material and Methods

The gathered imaterial consists first of the hearts
aind coronarv arteries of 403 captive wild vertebrates, that died in the Zoo of Antwerp or at the
capturing camiip of Epulu, and the aortas of 72 of
them (table 1). The anatomy as well as the pateney
of the coronary arterial tree of most of these anitoals was investig,ated by postmiortem arteriography.
A second group of imaterial includes the heart
anid coronary arteries of 144 vertebrates living in
freedom and killed in their natural habitat. The
aortas of 62 of them were
- also examined (table 2).
TIn both groups of animilals, the proximal segments of the main coronary trunks were removed
and fixed in formalin. Paraffin sections were prepared and stained with Masson's triehronme stain
(iron bematoxylin-ponceau-light green), and with
Weigert's resorein-fuehsin for the staining of
elastic tissue. Very often, Gabe's method (aldehydefuchsin) was used for the simultaneous staining
of elastic and connective tissue. We occasionallv
prepared frozen sections in order to stain lipids
with Sudan black B.
The partially dehydrated aortas were stained in
a saturated solution of Sudan IV in 70 per cent
ethyl alcohol for 10 minutes, and then differentiated in 70 per cent ethyl alcohol for 24 hours,
in order to remove excess stain. Frozen sections
were made in the stained areas.
Thirdly, in 88 of the captive anim-lals and in 51
of the free ones, blood samples were obtained by
venous or by cardiac puncture, and the following
determinations were made: lipidograim, total blood
cholesterol, cholesterol/phospholipid ratio, cholesterol fraction bound to the beta-lipoproteins, and,
in many cases, activity of the clearing factor, as
determnined by Hahn's technic.l4 Mfethods were described in a previous paper.15
Results
Degenerative Arterial Processes in Poikilothermic

843
Table 1

Captire Vertebrates
1. Reptiles: 38 specimens, ineludiiig represenitatives of
Cheloniidae (10)
Crocodylidae (4)
Varanidae (3)
Lacertidae (4)
Iguanidae (1)
Serpenits (16)
2. Birds: 123 specimeins, includinig represenitatives of
Stuthionifornies (2)
Galliformes (15)
Lariformes (3)
Balearicidae (8)
Sphenisciformes (2)
Anseriformes (27)
Cicoiiidae (30)
Falconidae (6)
Coraciiformes (20)
Passeriformes (10)
3. Mammals: 242 specimens, iincidiig
representatives of
Marsupialia (3)
Carnivores (28)
Pholidota (6)

Pininipedia (8)
Perissodactyla (8)
Suidae (6)
Ruminants (75)
Rodents (10)
Proboscidae (3)
Primates (95)

Table 2
Vertebrates in Freedom
1. Fishes: 40 specimens, iineludinig represenitatives of

Selachae (1)
Anguillidae (3)
Cyprinidae (20)
Thunnidae (10)
Istiophorus (6)
2. Bir ds: 36 specimens, ineludinig representatives of
Spheniscif ormes (29)
Ciconiidae (2)
Coraciif ormes (5)
3. AMaminals: 68 specimens, includinig
representatives of
Carnivores (3)
Suidae (35)
Ruminants (9)
Primates (21)

Animals

Elastosis of the initernal elastic membrane

of the coronary arteries was observed in some
sharks (I surus Nasus Donnaterre). In the involved arteries, elastosis was an isolated process, withlout any intimal sclerosis.
Among bony fishes, however, much more
Circulation, Volume XXVI, November

1962

characterized lesions were observed. This was

especially the case in the coronary arteries of
somne fishes (bluefin tuns: Thunnus Thynnius
L.). The first changes eonsisted in circular
or semiicircular nodules of hyperplastic intimiial tissue projecting into the vessel s luien

VA4
VrAESAEGER, )ElCOfIT()UTRT

8H44

Figure 2C
Abundi anlt lipid deposits in the thi kerned mail
coronary t? n/k of a immature Thnn?ns. Sudan

b1)/c/1

B X

(fig. 2A )

Figure 2A
Fat l intin4a/l 'cItctoau itt; 1/te rujtt c4't44t4r4 otertt1( {//' I t ttl}l
av
t1iif)4 1 (;7.I/i. I leifA
Iltsupc 'tX I. X ej,

2).5.

Tllw internal elatst i' mIlcbllltl)'al)

mi(lIerlleathi the inltial proliei-ations wa,s

so0li44mes rllptulle( aiid its frarinent s were
dlisJ)erse(l ill the hvyLerlplastie, intinia. Altflhou(g. fibroblasts ta-nd elolagyenois suhstaiiee
wxxeve the mi-laiin constituents of tle iollvlules, the
peseis(iiee of iinuerous histioeytes witlh tlearoxytopla4S1in -very often eonferre( to tihle lesiou-S
(lefillite tendencvy towatid atherosclerosis
(fi'. 213). Trplse (ealnge0s, elosel) reSelhl)liillg
thliose reee)itly (dmlesribed inI spawvning' Paciffic
Sablions bV Robertso. Wexlexr, alill Miller,'4
WClre0 found ill till of the' 10 illVtstlgated tlnlnielS rPlwey were xoun c, intiluiilt lure speleliliells
abmit :3 feet lo(,n, eatught in thle l)b-v of B'iscaIV
(Fainee) The main eoronary rtlerv of two
of thiem shiowe(d wilespreadl atherosclerosis
witlh abundant 4xtrial'T4ll1flar ljpid. d1e'posits

(fig. 2C).
(Certai llial)iallianu reptiles, iII (raptix'ity at
letlst, eal also (develop dlifflse ilntilall selerosis. This is tlhe (ease x ith eeritaiu opliidiains
of the typ)llo)s giolup: tlle coronary arteries
of a, largte ainaeoida (Eutoec.s marinaus L.)
A verp Zoo, slowevd fra"th rli
A
that died if
mneltation of thie intern'al. elastic( 111(lineb)ranc

t1igure zn
FJ"ca/ i14ti4441/l st'l'tr'osis i?/ tltt r'igh/4t c'r)4't0()ry',
ios !/ (4 1tioin/ fish. llt itJct-t X t).

arte'-

xvitlh slight fibr ous, tlhickeninlg of the iltimna.

Focal initiimial thickenlieng witlh ruiptured interutal elastic inenibrane was also fotunid in a
main coronary trunk of a 9-foot crocodile
(Cr'oco(dilats iloticts) living for mtiairany mnonithis
Circulation, Volum e X\X VI, Nov mbe r 19432

NATURAL HISTORY 1F ArlAHEROSCLEROSIS

in captivity at the Epulu camiip (Congcro). Ini

th1e lesiolns, abmndant listiocytes witli elear
cytoplasmn also sugg(-est a defilite ten(ledilr
towarl atherosc lerosis (fi(g. 3A aiid B). 1-ii
this animiiial, gross sills of coroiary s-clerosis
ws,ere see-n.
For fishes as well as for reptiles, the nmim
h)er of investigated specimelnes is too snall fotr
ain estimnationi of thle freqneiiev of arteriloslero(sis inl thlese (grolupsll of -verl tebrylates.
Frequency of Atherosclerosis among WarmBlooded Vertebrates

,ortie Atheoscletieosis

Siidanophilic fatty streaks of atlerosclrot' nature are relativelv ommll11on0 ill thIe
aorta of wvild mraImlInals ail(1 birds, and were
preseInt in nerl1y 30 per cent of tfle exallled
sIvecinleiis (table 3).
Aimioi cai)tive aniiiimals, the lipid (Iel)Osits
were very strik6ing ii thle aorta of a rlhea
(Rhcau IRotschild Brab.), of a l)eliean (1P%icU/ins Speick), of two wild boar (Su?s Scrafu
L.), of a 30-year-old eanmel (Ca}me/tus bactrianus LI.), of ani ai1dult biiffalo (Bubalus b)ubalis
L.) of a yak (Roephagrus grunWiens L.), of aii
oldI baboon (Pupio Paplo Desmarest), and(1 of
a youmig mnale ebiinpanzee (Pan Sch/iuvc
furtl/hii [Cigi])
Of the wild birds shot in freedom, nlolne(
shiowedl any sio'n of aortic atherosclertosis.
Verv slihlt lipid (leposits wvere observed i1n
thle aorta of eight of 35 wild boar (Sus Scnaofui

Bf

Figure 3
A. and 13. Focal intiwald sc lcrit in the caranarq
airteries of at. 6-foot cap/iet ecrocodul e. Thme 1??nt r0/Ifs hitioc
tqtcs With a ct atl? cyjotplasmnsShow cc dcj1Jit
anned Wheo
/h e lerrosis. WolViqed A,
il t e/
X 2.5; B, X 100.

i.) sihot ill theilr nIaltnlll'-

habitat. Of 1three'

(leer, ani adult feimale showed more

ICIispiciaius faitty- streaks, (dissemimlite(l ill its thoracic
aild abdomfinal aorta.

Table 3
4 ortic .4 therosclerosis
Captive specimens
Normal Atherosclerotic

1. Reptiles

2. Birds
3. Nlanimials
Marsupi.alia
Canilivores
Perissodactyla.
Suidae
Rumninriants
Rodents
Primates

Total
Cireulation, Volume AX VI, Nouvnilber

Free specimens

Normal

Atherosclerotic

Total

14

16

14

28
7

7
1

36

28

8
48
1962

13
24

10
47

15

38
13*4

846VATSESAE%JER,. DELCOJR T

846

Table 5
Inatimeal Sclerosis in Primates

Table 4
(o.o bus MonAeks
No.

I
4
9

4
)

Genus

C.
C.
C.
C.
C.
C.
C.
C.

Badius
Ba dius
Badius
Badius
Angolensis
Angolensis
Anigolensis
Aiigoleiisis

Sex

Aortic
atherosclerosis

F
F

F
F
Ml

++-

Amoong free primates, in lnonie of the fi-ve

and four Galago Demidovi [Fischer]) was

ani
sudaniophilic spot to be fo-und, in spite
of the fact that the serumii of one of the galagYos was turbid. Of the 11 African monhkeys.
two adult females of Colobus (Colobus auigolensis), a youngomale of Aseanius (Cercopithlascanius

Lemuridae
Callitrichidae aind Cebidae
Cereopithecidne
Pongidae

A:t

lIemnuridae (one Perodicticus Potto [Gmiiell

cts

With
intimal
sclerosis

[Audebert]),

and

an

old female

of Mangabey (Cercocebus aterrim us [Oudemnans]) showed widespread fatty streaks in

the thoracic aorta and less important ones in
the abdominal aorta (table 4).
Of the five examined ehinmpanzees shot in
freedom, two had localized atherosclerotic
lesions in the thoracie aorta: the lesions were
larger anid more important in a young adult
female than in a somewhat older adult muale.
Neither in eaptive nor in free aniiials were
aortie lesions complicated by caleification, by
imitramural hemorrhage, or by thronmbosis.
The aoirtie arch was rarelv affected alone, anid
it was rather commonier to find fatty patches
disseiminated along the deseending portion of
the thoracic aorta and stretching along the
abdominal portion. Such a dispersion of the
lesions was observed in birds as well as in
maiymals; in ungulata. at least, the thoracic
aorta seemed to be affected sooner thain the
abdomuinal portion.
Coronary Atherosclerosis

Coronary atheroselerosis is rare in our material; it was found in onlv nine of the 484
Nvarin-blooded vertebrates, i.e.. about 2 per
oeut of the examinedi speeimeiis. All of the
aiffected indiviiduals were captive: two were

Total

23
39
65

923_
515
401
125

vegetarian mammin-als (a mouffion, Ammotragus.

Lervia [Pallas], and a tapir, Tapirus Inlldictus
Desmarest); three were omnivorous mammrnals
(two chimnpanizees, Pani Sch weitfurthii [Gigl],
anid one little panda, Ailurus fudgen1s [Cuv.])
the other three were vegetarian birds (a
crownied crane, Balearica pavoninta (LA), and
three tree ducks, Dend-rocygn?a javanica

[Horf.].
Atherosclerosis of the nmouffloi 's coroniary
tree was very slight anid consisted exclusively
of snall sudanophilic spots in the in-tima s
diffuse thickeninig. More widespread in the
aniterior deseending braneh and in the eircumiflex braneh of the left coronary artery, were
the lesions of the tapir: they eonsisted not
onily of lipid deposits in the thickened fibrous
intimna but also of raised areas forminig real
atheromatous plaques (fig. 4).
The muore conspicuous lesions of the
chimupanzees were hunman-like atheromatous
plaques with foam cells and cholesterol crystals. In the little panida, however, they were
quite differenit: despite their diffuseness! and
the absence of atheroma, an oeclusive thromnbosis occurred in the anterior descending
branieh and killed the animal.17 In this case,
also, foam eells and cholesterol crvstals were
present in the thickened intimta.
Cholesterol crvstals were fouind onilv in the
coronary lesions of the omnivorous animals
(chimpanzees, panida). The lesion-is of the vegetarian vertebrates, either miaminlals (niioufflon,
tapir) or birds (crane, free (lucks). wTere quiite
free froni this sterol.
Diffuse intimnal sclerosis is a much. miiore
comnion finding thani atheroseler osis in the
coronarv arteries we investigated: it was ob'served in more than 25 per cent of the eaptivet
Circulation, Volumte XXVI, November 1962

NATI'RIAL 11FTOIIY ( ti' AIIElIiOSQLEIlO8ITS

8 47

.:-N

.1-

I.wlL..
I
.1

IT..
Figure 4
NItherloserotbe plaque in th e ri ht Ioronaryr(Jrer/f
of a tapir. Lipid infiltration owas pres(-nt in the
other coronary trunks of this animal. 1eigefrt:
X 10.

hinalll]Wds 1(i nearix 1 p rent ofdthfe

bir'1ds. il the ecaptiVe ver.teltrates of our11 innWterial, the freqnevey ()of liffinse intiual sclelSis se(1ems to be hlig.h.e.r amlong01' omlnil;vorouIs
andlherbi-vorous aininals than a.mong' earnruvannua.ls.
Amnong- primates, 52) per ecuit of whici slhowv
1 Of
t1mo0re Or less 1i1mportnIt; fibrolls thickeIing
the iuitina in their eoroniary arteries, the less
vu-lnierable to itimntal sclerosis wvere ltlelenorous

iidae, anid the mo-s Auileraievwere thwe polgidae (table 5).

Although1) nio sign. of coroarvy atherosclerosis couldl l)e diseoTered ini annualmIs shot in
-freedoi, the coronarr-v arteries of deer, roebuck, u ild boar, mnellys, anmd chimpanzees
often shiowved diffuLse intimlal selerosis, w-hiel
appqwarently j-intensifiedI wvith age. This is, howvever, very difficlLt to estimiiat e because the
vari.ations ini the vulnierabilitv to intimal
sclerosis are niot only specific. but also idividnal.
TI every case, ehimpaizees alI grorillas
(andl, to a less degree, orag-utans) are witlh
mi-an the onily species of thie atfnim-l kiigrdomi
in wic-leih dliftnse intfimal selerosis is a genleralize(d featuire antIi begins as ear ly as the first
weeks of life.
Of iiteiest iv as the d iseoverv of ani organizingo thiroimibus in the right eoronary artery
of a 7-month-old boar that was hunted and((irculation, Volatnoc XXVI, Norcmber 1.962

Figure n
Coronary thrombosis lin a 7-month-old :wild, boar.
[The thr-ombuzs deIclopedi in the righlt coronary!
ar-terq in spite of the absentce of atherosclerosis
in the coronitary arilter-ial tree a1wl( in1 tlhe aorta.
WcVigert: A, X 2.t5; B, X .10.

slolt. (fig. 5) :in thlis animial, throimibosis de1,Velopled on pire initimyial sclerosis, in spite of'
the v\lerified absence of any lipid deposit eith-elr
in the aorta or in t1e coroniary arterial tree.
Biochemical Results
Total Cholesterol anid Phospholipids

Thle level of totail cholesterol is by nio neanis

echaracteristie of the species. The range is
comiiprised betw-eeni 7.5 aIndi 250 nig. per ceniit
for., whole mnateriail. Triel.e are nio strikingy
n,genierie differences in. t lie clholesterol /plospliolipid iratio, amiong captilVe orl amllloln free
specimnes (old )looCled, 0.56 to 0.9; carntivorIols. 0.50 to 0.60( vegetariani ande onnfiv-or(Ins. 0.70 to 1.12). indilviduil variationis maxlie great (table 6).

VASTESAEGER, DELCOIRT

848
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44 P;,

8-49

NATURAL HISTORY OF ATHEROSCLEROSIS

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Circulation, Volume XXVI, November 1962

85Q''

VASTESAEGER, DELCOURT

-W

C i ccct. I.L 'a. t

CAAN,VOautL

PRI tATE4

Figure 6
Evolution of the lipidogram in different groups of
mammals. Arrow indicates: Left. Liptidographic
pattern in primates; (top) cercopithecidae, (center) galago (ga'lagidae%, (bottonm) potto (lorisidae.). Center. Lipidographic pattern in baboons;
in freedom (straight line), in captivity (dotted
line). Right. Lipidographic pattern in (top) ungulates, (center) carnivora, (bottom) pholidota.
Whole Lipoproteins

The lipidogram pattern is more characteristic of the species. As a rule, the lipidogram
seems to be related to evolution: the more
evoluted the species, the more the alpha-lipoprotein fraction becomes prominent (fig. 6).
Isolation and Characterization of Beta-Lipoproteins

Burstein"5 described a method suitable for

quantitative separation of beta-lipoproteins.
Dextran sulfate of high molecular weight and
a molar solution of caleium chloride are added
to the serum. Beta-lipoproteins are by this
way selectively precipitated. But even in human serum, there is a rather strict dependence
between the precipitation of beta-lipoproteins
and the molecular weight of dextran sulf ate.18
The procedure of Burstein remains questionable, however, when one tries to study lipoproteins in animals other than man.

Figure 7
Lipidograms of a wild boar: I. in freedom; II., in
captivity.

Lipidograms confirm that the beta-lipoproteins are correctly separated by this method,
in cold-blooded vertebrates as well as in mammals. However, Sakagami and Zilversmit,9
have shown that dextran sulfate is a suitable
reagent to separate quantitatively the betalipoproteins of the dog serum.
In our experience, the precipitation is not
linked with any antigenic property of lipoproteins. Using antihuman beta-lipoprotein
serum, we found fractions having a common
antigenicity with human fraction only in
monkeys and in some antelopes (Cephalophus
monticola [Thunb] ).
Generally speaking, as in human serum, the
percentage of cholesterol bound to beta-lipoproteins in wild animals was not very different from the percentage of this fraction as
demonstrated by lipidogram. But absolute
levels were quite dispersed. Extensive studies
should be done on many individuals in each
species before one may even attempt to systematize the biochemical results and the specific trend toward atherosclerosis (table 7).
Lipoproteins and Captivity

It can be seen, either by lipidograms or by

chemical determinations, that captivity has a
deleterious effect on the beta-lipoprotein levels. In lipidograms, the beta peak becomes
very prominent; the total cholesterol is much
higher, as is the level of cholesterol bound to
beta-lipoproteins (fig. 7, table 7). The changes
Circulation, Volume XXVI, November 1962

851

NATURAL HISTORY OF ATHEROSCLEROSIS

Table 7
Cholesterol and Phospholipid Levels and Ratios
Status

Species

Wild boar (Sus searofa) I1

2
3
Pangolins
(Manis tricuspis) 1
2

Mangabeys
(Cereocebus ateriimus) 1
2
3
Baboons*
(Papio doguera) 1
2
3

Total

Cholesterol
a
18

Phospholipids

,8 Phospholipids

%,

Total

18

13 Cholesterol

Freedom
Captivity, Europe
Captivity, Europe

71 51
114 104
193 139

20
10
54

28
9
27

176
156
144

84
119
100

92
37
44

4.6

Freedom
Captivity, Congo

145 110 35
220 97 133

24
60

256 218
243 160

38
83

1.08
.62

Captivity, Congo
Capitivity, Europe
Captivity, Europe

127
204
211

52 75
78 125
92 119

60
61
57

160 118 42
256 73 189
202 73 169

.56
1.46
1.08

175 26 149 85
214 100 114
Captivity, Congo
142 20 122 87
158 82 76
Captivity, Congo
144
Freedom
4
Freedom
157 58 99 63
212 118 94
170 113 57 33
5
Freedom
272 146 126
70 55 16 27
6
160 73 87
Freedom
*Tn spite of slight specific differences, our figures are in agreement with those of McGill and
found in Kenya baboons living in freedom, as well as with those of Van Zyl and Kerrick'
captive specimens of Papio ursinus.

in the phospholipids, either total or bound to

beta-lipoproteins, are less pronounced.
Discussion
Until now, most investigations concerning
spontaneous degenerative processes have been
carried out either on domestic animals or on
captive wild animals. Of course the domestic
ones have been deeply modified in their morphology as well as in their physiology by the
artificial selection that man has brought to
bear on them, sometimes during thousands of
years.

One must then ask if it is not human influence that, either by domestication or by
captivity, has favored the birth and the development of the arterial degenerative processes observed in the examined animals. In
order to answer this question, one should be
able to compare under rigorous conditions the
arterial state of animals of the same species,
captive and in their wild state. First of all,
in view of the density of human population
and its dispersion over all the lands of the
globe, and also of the multiplicity and power
of the commanding means of man over nature,
it is difficult to conceive that there could still
Circulation, Volume XXVI. November 1962

3.7
.81

.77
.62
.95
2.2
5.4
co-workers'0
obtained in

exist animals that entirely escape human influence. If such conditions still exist, they
are realized in the sea: the discovery of intimal sclerosis and of atherosclerosis in tunny
fishes caught in their natural habitat brings
proof that the two processes are not a consequence of captivity in the case of coldblooded vertebrates.
As far as warm-blooded animals are concerned, there are still wild specimens living
in places almost inaccessible to man and having only brief and incidental contacts with
him, that have kept to strictly natural foodstuffs and to a natural way of life. In order
to establish the influence of captivity on the
genesis and evolution of arteriosclerosis, there
still remains the difficulty of procuring individuals of the same race that are comparable
as far as age, sex, and stage of development
are concerned. All these conditions are far
from being fulfilled in our material: this is
why we cannot draw any firm conclusion
concerning the influence of captivity on the
incidence and evolution of intimal sclerosis
and atherosclerosis in birds and mammals.
Nevertheless, the lesions we observed in

852

deer, in colobi, and in chimpanzees living in

freedom bring evidence that, at least in ruminants and in primates, neither intimal sclerosis nor atherosclerosis is a mere consequence
of captivity. From the comparison of sections
made in the same coronary trunk of young
captive chimpanzees and of free chimpanzees
nearly of the same age, we got the impression
that the importance of individual variations
in the stage of intimal sclerosis in this species
may occasionally surpass the influence of captivity on this process.
Our biochemical results are also rather
disappointing for the study of the natural
history of atherosclerosis. Before the results
are discussed, the following points should be
emphasized. 1. Even in freedom, an animal
presumed normal is not necessarily normal.
Specimens shot or trapped in their habitat
are usually disabled or ill, the healthy and
fit ones escaping more easily. In any case, we
had the opportunity to see how severely tropical animals are infested with parasites (table
6); unfortunately the influence of parasites
on the lipoproteins pattern is still unknown.
2. Captive animals are not normal. Two kinds
of captivity are to be considered: in the first
one, the animals remain in their natural climate, not far away from their original habitat; in the second one, the animals are sent
to zoos in far distant countries. In the first
case their natural feeding grounds are little
disturbed, but in the second case diet and
environmental conditions are quite different.
As compared with the values recorded in
freedom, both types of captivity significantly
alter lipoproteins as well as cholesterol levels.
This was observed not only in European suidae and carnivora, but also in African ungulates and primates. This deleterious effect
of captivity is impressively confirmed by a
previously published study of 60 young chimpanzees living for many months at the medical
Laboratory of Stanleyville.22 Table 8 summarizes the results of this study. The total
blood cholesterol ranged between 120 and 470
mg. per cent, the mean value being 250 mg.
per cent. In all specimens the cholesterol

VASTESAEGER, DELCOURT
fraction bound to the beta-lipoproteins was
quite large. No relationship between cholesterol level and age, sex, or length of captivity
could be established.
In any case, our biochemical results bring
accumulated evidence that captivity may significantly alter some aspects of the lipidic
metabolism in animals. This should be kept
in mind when one seeks suitable material for
experimental atherosclerosis.
As far as comparative arterial pathology
is concerned, our observations confirm those
of Fox dealing with the widely separated
varieties of the affected vertebrates. They also
confirm the statement, at first made by Fox,
that the lesions are mainly distributed in the
aorta, the great majority of the involved
specimens showing this distribution alone.
Our results disagree, however, with those of
Fox concerning frequency and specific incidence of atherosclerosis: in Fox 's material
"lipidic deposits were exceedingly rare," in
ours, nearly 30 per cent of the examined aortas were affected. On the other hand, in Fox's
material, primates were less vulnerable than
carnivora and ungulates to degenerative arterial disease: on the contrary, it was among
primates that we found the highest percentage
of aortic lesions. Such a discrepancy can of
course be explained by the; difference of conditions of captivity in the zoos of Antwerp
and Philadelphia.
Different local conditions of diet are probably not the only factors responsible for the
discrepaney between Fox's figures and ours:
in the same Zoo of Philadelphia, between 1931
and 1956, Ratcliffe and Cronin7 observed an
inereasing frequency of arteriosclerosis in
captive wild animals, independent of age, and
diet.
The results of our investigations are very
difficult to compare with the figures published
by Rateliffe and Cronin. As a matter of fact,
the criteria of these authors are quite different from ours: their practice has been "to
ignore isolated plaques of the aorta and larger
branches, unless they were accompanied by
vascular disease of the heart, kidney or
Circulation, Volume XXVI, November 196.t

853

NATURAL IIISTORtY OF ATIJEROSULEROSIS

sptleen.' Furithermiore. tlhey imla-de 110 elihe(late effort to separate atherosclerosis from
otler formus of arterioselerosis.
The numiber of our specimenes is, of course,
too small to allowv even a ronghi estimiiate of
the frequenev of both intiinal sclerosis and
atheroselerosis in eaeli speeies of vertebrates.
But7 as far as the incidence of spontaneous
atheroselerosis is concerned, w-e have been
struck by its greater frequlency amiong certamn species stueh as tree duekls (three of five
aninnials had eoronary atherosclerosis) and
amnono,g cenera like cercopithecidae anid al)es.
The frequeney of atheroselerosis is hig,her in
the prima-tes thani in the ungrulates, and hiigher
in the ungulates than in the carnivora.
On tile othe-r lhand, in its hIman incidence
diffuise intinial selerosis appears as a generalize(1 process, beginnling as soonl as the first
Aveeks of life in white as wcll as in Bantu
people?3 Such universal preeoeity among h-i
111811 beiniis gave manv autlhors the imnpressioni
that diffuise intinual selerosis could be a physiologic proeess, growingf progressively with
advancingf agre. AAWhereas intimal seletosis is
foullnd. in all voungr human aduilts, this is far
fi-oun the ease in other species.
-Although the facts brought about by our11
iinvYesticgations are very fragmentary, tley are
qutiite sufficienit to establishl that not onilv the
ineidenee of diffuse intimal sclerosis but also
the age at which it appears vary eonisiderablv
from one speeies to another. Obviously, the
fact that we cannot know the real agfe of captive animals makes any kind of precision
difficult, especially if the variable life span
of the different species is taken inito account.
Nevertheless, the faet that the coronary arteries of a captive wild pig, which died of
old age (fig. 8, top), show hardly ml-ore advaniced lesions than those of a 5-year-old
gorilla (fig. 8, bottom) proves undeniably the
existence of specific differences in the precocity of the fibrous thickeniing of the coroaiavy
intima. We could multiply suci examiples Cand
show that of all mrlammals the anthropoid apes
and mrlanl are the only ones to present systemnCirculation, Volutme XXVI, November

1962

Figure 8

'1'oj. Anterior descending branch of an o1(1 cO]pti

wart-hog. The internal elastic

ne mbran

iv

stretched and ruptured, bat thte jatiima is moloeiEately thickened. lVeigert X 40. Bottoml. Anterior
desceniding branch of a yloun ignInmature fecna7
gorilla. Tfhe internal elastic membrane is as thick
as that in the wart-hog.

atically importanit fibrous changtes of the coronary iintima before puberty.

The larger number of primnates we could
investigfate gave us the impression that in this
or(ler at least the frequency and the precocity
of initiial fibrosis increase with the degree of
evolution (table 5).
In ouir experienee, spontaneous atherosecierosis always develops in arteries alreadly impaired by iintimal selerosis. In otlher vwords,
iintimal sclerosis appears as a basic prelimilmary lCondition for spontanieous atherogenesis.
To what extent alterations of lipidi netabolism facilitate spontaneous atherosilerosis ini
thie fibirous initiina is still unikniowin, btut it is

firllliy established tihat atherosclerosis

may

854

spontaneously develop in animals having quite

different diets, and especially in fishes feeding on a diet rich in unsaturated fats; in
captive birds whose dietary fats are mostly
unsaturated (crane, tree duck, pelican) ; in
wild mammals whose diet is free of animal
fats, in freedom as well as in captivity (deer,
camel, tapir).
These facts indicate that there is no absolute and uniform relationship-i.e., applicable
to every vertebrate species and to every individual in a given species between the origin
or the chemical nature of dietary fats and
the development of atherosclerosis. Inbred
specific and individual differences in vulnerability to intimal sclerosis, reinforced by
specific and individual differences in enzymatic activity of lipid metabolism, yield a
better explanation of the great variability in
the incidence of atheroselerosis in different
species and in different individuals of the
same species. Captivity or way of life (diet,
physical activity, and stress) are further variable elements in the problem of atherogenesis
in wild animals.
Summary
In order to complete the fragmentary
knowledge of the natural history of atherosclerosis, the authors examined the coronary
arteries of 403 captive and 146 wild vertebrates living in freedom, as well as the aortas
of 134 of them. In 88 of the captive animals
and in 51 of the free individuals, lipidogram,
total cholesterol, beta-cholesterol and cholesterol/phospholipid ratio were also determined.
In this material, spontaileous atherosclerosis
always developed in arteries already impaired
by intimal selerosis, and intimal sclerosis
seems to be a basic condition for spontaneous
atherogenesis.
Lipid deposits were found in the intima of
29 per cent of the examined aortas, but coronary atherosclerosis was present in only 2
per cent of the captive specimens. The discovery of coronary atherosclerosis in free
tunny fishes, caught in the Bay of Biscay,
brings proof that atheroselerosis is neither

VASTESAEGER, DELCOURT
the privilege of warm-blooded vertebrates nor
a consequence of captivity.
Unfortunately, the material collected does
nlot permit even a rough estimate of the influence of captivity on the development of
atherosclerosis. It was observed in any case
that captivity significantly alters some aspects
of the lipidic metabolismi in animals, and
especially the beta-lipoprotein levels as well
as the beta-cholesterol fraction.
Spontaneous atherosclerosis may develop in
fishes living in their natural habitat, feeding
on a diet rich in unsaturated fats; in captive
birds whose dietary fats are mostly unsaturated (crane, tree duck, pelican) ; and in
wild mammals whose diet is free of animal
fats, in freedom as well as in captivity (deer,
eamel, tapir).
References
1. LOBSTEIN, J. G. C. F. M.: Traite d 'anatomie
pathologique. Paris, Levrault, 1833.
2. M6NCKEBERG, J. G.: t;ber die reine Mediaverkalkung der Extreiitiitenarterien urnd ihr Verhalten zur Arteriosklerose. Virchows Arch. f.
path. Anat. 171: 141, 1903.
3. World Health Organization: Rapport d 'un groupe
d 'etude: Classification des lesions d 'atheroselerose. Palais des Nations, Geneve, 1958.
4. MARCHAND, F.: uber Arterioskierose (AtheroSklerose). Verhandl. (T. 21. Kong. F. inn. Med.
21: 23, 1904.
5. Fox, H.: Disease in Captive Wild Mammals
and Birds. Philadelphia, J. B. Lippincott Company, 1923.
6. Fox, H.: In Cowdry, E. V.: Arteriosclerosis,
a Survey of the Problem. New York, The
Macmillan Company, 1933, p. 153.
7. RATCLIFFE, H. L., AND CRONIN, 1. T.: Changing
frequency of arteriosclerosis in mammals and
birds at the Philadelphia Zoological Garden.
Circulation 18: 41, 1958.
8. RATCLIFFE, H. L., YERASIMIDEs, T. G., AND
ELLIOTT, G. A.: Changes in the character and
location of arterial lesions in mammals and
birds in the Philadelphia Zoological Garden.
Circulation 21: 730, 1960.
9. DAUBER, D. V.: Spontaneous arteriosclerosis in
chickens. Arch. Path. 38: 46, 1944.
10. BRAGDON, J. H.: Spontaneous atherosclerosis in
the rabbit. Circulation 5: 641, 1952.
11. LINDSAY, S., CHAIKoFF, I. L., AND GILMORE,
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NATURAL HISTORY OF ATHEROSCLEROSIS

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Men of science who mean to embrace the principles of the experimental method as a
whole, must fulfill two classes of conditions and must possess two qualities of mind
which are indispensable if they are to reach their goal and succeed in the discovery of
truth. First, they must have ideas which they submit to the control of facts; but at
thie same time they must make sure that the facts which serve as starting point or as
control for the idea are correct and well established; they must be at once observers
and experimenters.-CLJAUDE BERNARD. An Introduction to the Study of Experimental
Medicine. New York, The Macmillan Company, 1927, p. 21.

Circukation, Volume XXVI, November 1962