THERIOGENOLOGY

CLINICAL, MORPHOLOGICAL AND ENDOCRINOLOGICAL

ASPECTS OF CRYPTORCHIDISM IN THE HORSE.

Coryn, M., De Moor, A.W, Bouters, R. and

Vandeplassche, M.

Faculty of Veterinary Medicine, Department

of Reproduction and Obstetrics,
State University of Ghent,
Casinoplein 24, B-9000 Gent (Belgium)
H Department of Surgery.

Received for publication: April 14, 1981.

Accepted: September 8, 1981.

SUMMARY

The authors analysed clinical, histological and hormo-

nal data obtained from 205 cryptorchid horses. The majori-
ty of the unilaterally and bilaterally retained testes were
located in the inguinal canal; however, the ratio of ingui-
naL vs abdominal retention appeared to decrease with advan-
cing age. In unilateral cryptorchidism, a pronounced pre-
ference was noted for left abdominal retention, whereas for
inguinal cryptorchids, the retained testes occurred equally
on both sides. Right inguinal retention was found to de-
crease with advancing age. Histology of cryptorchid testes
revealed apparently normal Leydig cells and arrested sper-
matogenesis. PLasma testosterone concentrations were simi-
lar in normal stallions and unilateral cryptorchids, even
in those which had the scrotal testis removed. Plasma
oestradiol-17S levels were lower in unilateral cryptor-
chids than in stallions.

The authors wish to thank Mr. R. Verbeke and Mr. D. Ver-

eecke for their technical assistance.

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THERIOGENOLOGY

INTRODUCTION

The incidence according to side (right or left) and

location of equine cryptorchid testes have been the subject
of several reports, but in only few of the investigations
was the number of animals sufficient to allow statistical
analysis (1, 2, 3). Moreover, only one of these reports
deals with the influence of breed and age upon these pheno-
mena (2). Comparative studies of steroid plasma levels in
stallions, geldings and cryptorchids are also rare (4, 5).
The present report is the result of the clinical, morpho-
logical and endocrinological investigation of cryptorchid
stallions.

MATERIAL AND METHODS

The subject of this study were 205 cryptorchid stal-

Lions presented for castration at the surgical clinic du-
ring the period 1970-1980. These cryptorchids were of va-
rious breeds and were aged between 1 and 5 years, with the
majority (71%) aged between 1 and 3 years. They had one or
both testes retained either inguinally, abdominally or ab-
domino-inguinally. Inguinal testes were situated between
the internal inguinal ring and the scrotum, abdominal tes-
tes were retained in the abdominal cavity and abdomino-in-
guinal testes were in the abdomen with the epididymal tail
and a Loop of the deferent duct, or both, present in the
inguinal canal.
The position of the non-scrotal testis was diagnosed
by palpation of the inguinal region, or by rectal palpation
on the standing or anaesthetized horse. In some cases, it
was difficult to distinguish a uni- or bilateral cryptor-
chid from a castrated or unilaterally castrated horse.
Frequently cases were seen in which only one, probably
scrotal, testis had been removed during a previous attempt
to castrate the horse. An inguinal testis that was not
found during this previous operation may eventually des-
cend Later on; the horse is then wrongLy presented as a
unilateral cryptorchid.
In most cases inwhichatestis has been removed, a "castra-
tion" scar can be seen, especially on the anaesthetized
horse. This scar is characterized by a variable degree of
retraction of the skin, due to adhesions of the skin and
subcutaneous tissue to the vaginal process and spermatic
cord. A Linear scar of the skin and subcutis is not proof
of a removaL of the testis on this side. It is indeed pos-
sible that a skin incision has been made and even an ingui-
nal exploration performed but that no testis had been found.
After uni- or bilateral castration the deferent duct is

490 OCTOBER 1981VOL.16N0.4

 THERIOGENOLOGY

always found at rectal exploration to enter the internal

inguinal ring on the side of the castration. However in
horses castrated at a very young age the deferent duct is
not easily recognized at a later age and can be confused
with the gubernaculum testis. In abdominal cryptorchidism,
a small gubernaculum and vaginal process but not the defe-
rent duct can be palpated rectally. In 37 cases of abdo-
minal cryptorchidism examined by one surgeon (x) the testis
could be palpated with certainty in 24 animals and presumed
to have been palpated in 6. From 8 horses with abdomino-
inguinal cryptorchidism the testis could be palpated rec-
tally in 5 cases. Rectal palpation was done on the stan-
ding horse after an intravenous injection of atropine fol-
lowed by xylazine at respective doses of 0.01 mg/kg and
0.8 mg/kg. In untractable horses where a thorough rectal
exploration even after xylazine premeditation proved dan-
gerous the palpation was done on the horse in lateral re-
cumbency after induction of anesthesia with guaifenesin
(glyceryl guaiacolate) at a dose of 0.1 g/kg followed by
haLothane/N20/02 in a closed anesthesia circuit.
In clinically doubtful cases a testosterone and oestrogen
response to an injection of HCG or even an inguinal sur-
gical exploration can be performed.
Blood samples from the jugular vein were collected in-
to heparinized tubes between 10.00 and 12.00 a.m. After
centrifugation the plasma was stored at -2O'C until assay-
ed. The concentration of testosterone and oestradiol-13$
was measured by radioimmunoassay. Plasma samples of 15
normal stallions and 15 geldings were included in the as-
say. The testosterone antiserum, raised in rabbits against
testosterone-3-(0-carboxy) methyloxime-BSA, cross-reacted
with Sa-dihydrotestosterone (28.0"/,),5a-androstane-3a ,
L78-diol (5.0%), n4-androstenedione (0.3%) and n5-andro-
stenediol (0.5%). A suitable standard curve (range : LO-
200 pg testosterone) was obtained with 0.1 ml antiserum
(l:lO.OOO), binding 60% of the added (1,2,6,7-3H)-testo-
sterone (0.1 ml, 53.000 dpm). Testosterone, added to 0.1
ml plasma, was recovered for almost 100% over the whole
range of the assay. For plasma samples, the mean within-
and between-assay precision was 9% (coefficient of varia-
tion). The oestradiol-L7B antiserum, raised in sheep a-
gainst 176 -oestradiol-3-hemisuccinate-BSA, cross-reacted
with oestrone (9.2%), oestriol (2.3%) and oestradiol-17e
(0.9%). A suitable standard curve (range : 5-320 pg oes-
tradiol-17e ) was obtained with 0.5 ml antiserum (1:60.000),
binding 40% of the added (2,4,6,7-3H)-oestradiol-L7@ (0.1
ml, 21.000 dpm). When known amounts of oestradiol-176 ,
added to 1 ml plasma, were assayed a regression line with
the equation y= 19 + 0.85 x (y=pg obtained, x=pg added)
was obtained. The precision of the assay was determined

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by the repeated assay of different plasma samples (1 ml) :

124 + 6 pg (Y + S-D., n= 9), LO4 + 4 pg, 74 + 2 pg, 57 +
3 Pg, 23 _+ 2 pg. NO chromatographic steps w&e include;
in the assays.
The cryptorchid testis, and eventually the contrala-
teral descended testis, were weighed and 5 gr of testicu-
lar parenchyma were fixed in formaldehyde. Paraffin sec-
tions (51-1thick) were stained with haematoxylin and eosin.
Statistical analysis was carried out with a X2-test (cli-
nical results) or a "Student's" t-test (morphological and
endocrinological results), using the method described by
Snedecor and Cochran (1967).

RESULTS

Clinical : In a total of 205 cryptorchid stallions 187

(91.2%) were found to be affected unilaterally and 18
(8.8%) bilaterally. Of 187 unilaterally retained testes
126 (67.4%) were situated in the inguinal canal, 51 (27.3%)
in the abdominal cavity and LO (5.3%) were abdomino-ingui-
nal (Ping abd <0.005). In ponies the respective num-
bers and pkrcentages were : 21 (70-O%), 8 (26.7%) and L
(3.3%) (Pin ,abd.' 0.05). When unilateral cryptorchids
.z:
were classi ied by age and position of the retained tes-
tis, the percentage of inguinal testes decreased with ad-
vancing age : 82.6% (38/46) between 1 and 2 years, 72-G%
(54/75) between 2 and 3 and 61.7% (29/47) between 3 and
5 years. Of the 36 bilaterally retained testes 24 (66.7%)
were Located inquinallv, LO (27.8%) abdominallv and 2
(5.5%) abdominolinguinaily (Ping.,abd_ = 0.05): Of the 18
bilaterally affected stallions 11 (61.1%) had two inguinal
testes and 4 (22.2%) two abdominal testes, L (5.6%) an in-
guinal and an abdominal testis, 1 (5.6%) an inguinal and
an abdomino-inguinal testis and 1 (5.6%) an abdominal and
an abdomino-inguinal testis.
The overall incidence of unilateral cryptorchidism was
slightly, but not significantly, higher for the Left side:
LOO (53.5%) : 87 (46.5%). For ponies a ratio of 13 (44.8%) :
16 (55.2%) was found. There was a tendency although not
significant, for right inquinal cryptorchidism : 68 (54.0%):
58 (46-C%), and a significant higher frequency (P < 0.005)
of Left abdominal cryptorchidism : 38 (74.5%) : 13 ,(25.5%).
For ponies the ratios were respectively : 15 (71.4%) : 6
(28_5%)(P <0.25) and 7 (87.5%) : 1 (12.5%) (P=O.L) - ClaSSi-
fication of unilateral inguinal cryptorchids by age and side
of retention revealed a decreasing percentage of retained
right testes with advancing age : 63.2% (24/38) between L
and 2 years, 55.6% (30/54) between 2 and 3 and 34.5% (10/29)
between 3 and 5year.s.

492 OCTOBER 1981 VOL. 16 NO. 4

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Morpholoqical: No significant difference in weight was

found between right and Left retained inguinal"ar abdomi-
nal testes : Ing.right : 35 + 26 g (x + S-D., n=L9), ing.
Left : 37 + 22 g (n=L8), abdTright : 47 + 14 g (n=4), abd.
Left : 36 7 17 g (n=9). For the corresponding scrotal tes-
tes no significant difference in weight was noted between
sides : Right : 204 _+ 55 g (n=L6), left : 183 + 51 g (n=L8).
No ponies nor cryptorchids younger than two years were in-
cluded in this comparison. In inguinal and abdominal tes-
tes, normal Leydig cells were found in the interstitial tis-
sue. The diameter of the seminiferous tubules was only half
of that in scrotal testes : 83 + 30 (n=35) vs 152 + 30 (n=35),
and of the same magnitude in inguinal and abdominal testes.
Moreover spermatogenesis did not proceed beyond the A- or
B-spermatogonia in abdominal testes, or young primary sper-
matocytes in inguinal testes.

Endocrinoloqical: No significant difference in plasma tes-

tosterone Levels was noted between stallions (0.71 + 0.46
ng/ml, n=L5) and unilateral cryptorchids with an intact
scrotal testis (0.62 + 0.41 ng/mL, n=74). No difference
was seen between ingulnal and abdominal unilateral cryp-
torchids, respectively 0.65 + 0.42 ng/mL (n=58) and 0.52
+ 0.35 ng/mL (n=L6). Even in unilateral cryptorchids in
Which the scrotal testis had been removed previously, tes-
tosterone concentrations (0.76 + 0.46 ng/mL, n=9) were com-
parable to those in stallions. -Basal concentrations in
geldings were 0.19 + 0.09 ng/ml (n=L5). Oestradiol-L7B
plasma concentrations in unilateral cryptorchids with an
intact scrotal testis (34 + 21 pg/mL, n=39) were Lower
(P=O.Ol) than in stallions-(65 + 34 pg/mL, n=L2) and very
significantly higher (P <0.005)-than the basal Levels in
geldings (15 f 6 pg/ml, n=7).

DISCUSSION

Inguinal cryptorchidism can usually be diagnosed cli-

nically on the standing sedated horse-by a thorough and
high inguinal palpation except in cases in which the in-
guinal testis is Located close to the internal inguinal
ring. In uni- or bilaterally castrated horses indistinct
"castration scars" usually cannot be palpated on the stan-
ding animal: inspection and palpation of the external in-
guinal region have then to be done on the anesthetised
animal. If a paramedian or flank approach are used for
the removal of an abdominal testis it is absolutely neces-
sary to differentiate between inguinal and abdominal cryp-
torchidism. This differentiation usually can be made by
rectal palpation on the standing or anesthetised horse.
Steroid analysis is indicated when no testis can be pal-

OCTOBER 1981 VOL. 16 NO. 4 493

 THEFUOGENOLOGY

pated either by inguinal or rectal palpation and no dis-

tinct"castration scarsll together with a deferent duct are
found. Steroid analysis is also useful when general anes-
thesia is not indicated in horses that are very difficult
to examine or in rare horses that retain a stallion beha-
viour after castration or after one or more unsuccessful
attempts at castration.
Bilateral retention was found in 9% of the horses pre-
sented for cryptorchidectomy. Similar (1, 2, 6) or higher
percentages (up to k8%) (3, 7, 8) have been reported in the
Literature. Differences could be due to different number
of animals unilaterally castrated prior to admisbion.
Of the unilateral cryptorchids the majority (67%) had
the testis located in the inquinal canal. This is in qood
agreement with the findings of most investigators (2, 6,
7, 8) (range : 61 to 68%), but in contrast to reports by
others (1, 3)‘ 39 and 43% respectively. In ponies, the
preference for inguinal retention was Less pronounced. In
bilateral cryptorchids the majority (67%) of the testes
also were retained inguinally, as reported by some authors
(6, 8) (70%, 63%). The same authors mentioned above (L,3)
again found a minority of inguinal testes (40%, 33%). Of
the bilateral cryptorchids 83% had the two testes either
in the inguinal canal or in the abdominal cavity. ALL per-
centages reported in Literature exceed 80% (1, 2, 3, 6, 7,
8).
The ratio of inguinal to abdominal retention was found to
decrease markedly with advancing age. This supports the
findings of Cox (1979) and demonstrates that descent of
an inguinal testis may occur at Later age. Testicular
descent is normally completed at Last a few weeks after
birth (9).
Consequently the percentage of inguinal retention for
a group of cryptorchids depends on age and pbssibly breed
distribution, which could explain most of the contradiction
in Literature.
The overall incidence of unilateral cryptorchidism was
somewhat, but not significantly, higher for the Left side.
Some investigators (1, 2, 3) note a slight tendency for
Left, others (6, 8) for right retention. Cox (1979) found
that ponies had significantly (P 10.005) more right re-
tained testes. This could not be concluded from our ma-
terial although a tendency in that direction was noted.
Of the unilateral abdominal cryptorchids 75% (P eO.005)
had the testis retained at the Left side. A higher fre-
quency of Left abdominal retention is generally accepted
(1, 2, 3, 8, LO) and could be due to the fact that, at the
time of normal testicular descent, the regression of the
hypertrophied foetal testes is slower at the left side
(11). For ponies however, the preference for left sided

494 OCTOBER 1981 VOL. 16 NO. 4

 THERIOGENOLOGY

abdominal retention was not so clear (P=O.l). cox (1979)

even noted that less than half of the abdominal cryptor-
chid ponies (55/123) were affected on the Left side. Of
the unilateral inquinal cryptorchids, the retained testes
were equally distributedonbothsides. This confirms the
findings of several investigators (1, 2, 6, LO). Others
(3, 8, 12) however, found a significant difference between
sides, aproximately 70% of the animals had the inguinal
testis on the right. In a group of ponies we noted that
right inguinal retention was more frequent. This was also
reported by Cox (1979) who noted moreover that this pre-
ference decreased with advancing age, resulting in an equal
distribution on both sides. We found that for larger
breeds most of the inguinal testes were retained at the
right side for animals between 1 and 2 years. For animals
between 3 and 5 years the opposite was noted. Hence, the
already mentioned late descent of inguinal testes obvious-
ly occurs mainly at the right side. From our findings and
these of Cox (1979) it becomes clear that for a group of
unilateral inguinal cryptorchids the side distribution of
the retained testes depends on breed and age.
Histological findings in inguinally and abdominally re-
tained testes were in agreement with those mentioned in Li-
terature: apparently normal Leydig cells develop in the
interstitial tissue (8). The seminiferous tubules are smal-
ler and there is only a single layer of spermatogonia and
Sertoli cells (7, 8). In inguinal testes however, there may
be a second layer of primary spermatocytes (7).
No difference in plasma testosterone Levels was noted
between normal stallions and unilateral cryptorchids, while
Levels in geldings were significantly Lower. This confirms
the findings of Ganjam and Kenney (1975). The absolute va-
lues found were considerably Lower than those obtained by
the authors mentioned, which can be explained by the fact
that our antiserum showed Less cross-reactivity.with DHT.
The fadt that plasma testosterone concentrations were the
same in unilateral cryptorchids of which the scrotal tes-
tis had been previously removed and in normal stallions
strongly suggests that the retained testis has a compen-
satory hypersecretion. A similar explanation has been gi-
ven for the observation that hemicastrates could not be
differentiated endocrinologically from animals with two
testes (5). Oestradiol-17fl plasma Levels were found to be
lower in unilateral cryptorchids than in normal stallions;
dissenting results had been reported (4).

REFERENCES

1. STANIC, M.N. Castration of cryptorchids. Mod. Vet.

Pratt., & (241, 30-33,(1960).

OCTOBER 1981 VOL. 16 NO. 4 495

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2. COX, J.E., EDWARDS, G.B. and NEAL, P.A. An analysis

of 500 cases of equine cryptorchidism. Eq. Vet. J.,
LL (2), LL3-116,(1979).

3. STICKLE, R.L. and FESSLER, J.F. Retrospective study of

350 cases of equine cryptorchidism. J. Am. Vet. Med.
Assoc., 172 (3), 343-346,(1978).

4. GANJAM, V.K. and KENNEY, R-M. Androgens and oestrogens

in normal and cryptorchid stallions. J. Reprod. Fert.,
Suppl. 23, 67-73,(1975).

5. COX, J.E., WILLIAMS, J-H., ROWE, P.H. and SMITH, J.A.

Testosterone in normal,cryptorchid and castrated male
horses. Eq. Vet. J., 5 (2), 85-90. (1973).

6. SCHEBITZ, H. and SOLLER, D. Zur Kastration des

Kryptorchiden. Berl. Miinch. Tier?kztL. Wschr., 93, LOL-
104,~(1980).

7. ARTHUR, G.H. The surgery of the equinecryptorchid. Vet.

Rec., 7-3, (16), 385-389,(1961).

8. BISHOP, M.W.H., DAVID, J.S.E. and MESSERVY, A. Cryptor-

chidism in the stallion. Proc. RoyaL Sot. Med., 59(8),
769-770, (1966).

9. BERGIN, w-c., GIER, H.T., MARION, G.B. and COFFMAN, J-R.

A developmental concept of equine cryptorchidism.
BioL. Reprod., 3, 82-92, (1970).

LO. SILBERSEIPE, E. Einiges iiber den Kryptorchidismus der

Pferde. Berl. Miinch. Tier%zztL. Wschr., 53, 432-434,
(1937).

11. SMITH, J.A. The development and descent of the testis in

the horse. Vet. Ann., u, 156,(1975).

12. SMITH, J.A. Studies on cryptorchidism in the horse. PH.

D.Thesis for Univ. of BristoL,(L973).

496 OCTOBER 1981 VOL. 16 NO. 4