A Klinefelter bull with a 1;29 translocation born to a fertile 61,XXX cow

BRIEF COMMUNICATIONS
COMMUNICATIONS BREVES
A Klinefelter bull with a 1;29 translocation born
to a fertile 61,XXX cow
Klinefelter's syndrome is caused by a chromosomal
anomaly in which individuals typically possess two
X chromosomes and a Y chromosome. This syndrome
has previously been reported in cattle (1-3). The chromosomal
anomaly is generally thought to be the result
of nondisjunction during meiosis in one of the parents.
Generally, most mammals with Klinefelter's syndrome
are expected to be phenotypically male and infertile.
Libido and service behavior may be completely normal
(3); however, ejaculates are azoospermic.
The bull described in this report was discovered
during a study involving the karyotyping of a complete
herd of purebred Charolais (4). He was born on
April 6, 1990 to a three-year-old dam, as the result of her
first breeding at two years of age by natural service. In
the fall of 1990, he was taken to the university's feedlot
for finishing. He weighed 786 kg (1730 lb) and was
142 cm tall at the shoulder at 25 months of age. His scrotal
circumference was 19 cm at 13 months of age and
18 cm at 25 months of age, compared to an average of
33.1 cm and 36.3 cm for one and two-year old Charolais
bulls, respectively (5). The length of the extended penis
at electroejaculation measured 24 cm, and the penis
appeared to be of normal size and conformation. The urethra,
prostate gland, and vesicular glands were felt to be
in the normal size range, as determined by transrectal palpation.
A seminal fluid sample collected by electroejaculation
contained no spermatozoa.
Blood was aseptically drawn by venipuncture, and a
culture of lymphocytes was established on the day of collection
with 0.5 mL of blood in 10 mL of Ham's FIO
medium, containing 15% fetal calf serum and 0.2 mL of
phytohemaglutinin (Wellmark Diagnostics, Guelph,
Ontario). A small skin sample was surgically taken
under local anesthetic, and fibroblast cultures were
established (6). Samples of myocardium, pericardium,
liver, kidney, and skeletal muscle were collected at
slaughter, and cell cultures were similarly established
from these tissues. Cell harvest, slide preparation, and
photomicroscopy for karyotyping was done in the usual
manner (6). Trypsin G banding (7) was done on the
Can Vet J 1994; 35: 182-184
Department of Animal and Poultry Science (Schmutz, Moker)
and Department of Herd Medicine and Theriogenology (Barth),
University of Saskatchewan, Saskatoon, Saskatchewan
S7N OWO.
182
Sheila M. Schmutz, Albert D. Barth, Jane S. Moker
x-X- ----
0 ~~~, f1 P ft ^ ;";0"X
Figure 1. Trypsin G banded karyotype of a Charolais bull,
60,XXY,t( 1 ;29).
slide preparations from lymphocytes. A karyotype of
60,XXY,t(1;29) (Figure 1) was found in preparations
from all lymphocytes and all fibroblasts examined from
skin, heart muscle, pericardium, liver, kidney, and
skeletal muscle.
Blood samples for serum testosterone analysis were
taken from an indwelling catheter in the external jugular
vein every 30 min over a six-hour period. This was
followed immediately by a gonadotropin releasing hormone
(GnRH) stimulation test, when 250 jig of GnRH
(Factrel, Ayerst Laboratories, Montreal, Quebec) were
injected intravenously and blood samples were taken
every 30 min for a two-hour period. Serum was harvested
from the blood samples and frozen for testosterone
assay at a later date. Testosterone was measured using
a modification of a radioimmunoassay (8); assay standards
were prepared in charcoal-stripped serum (0.02 ng/mL
to 10 ng/mL) and extracted with ether before assay,
rather than being put directly into assay tubes. Fifty
microliter aliquots (instead of 200 ,uL) of standards
and samples were extracted in 2 mL diethyl ether. The
sensitivity of the assay was 0.02 ng/mL or 0.07 nmol/L.
When 1.0 ng or 2.5 ng of testosterone were added to
I mL of serum, the values obtained after correction
for endogenous testosterone were 3.75 ± 0.52 nmol/L and
8.74 ± 0.97 nmol/L (x ± SEM), respectively. All samples
were analyzed in a single assay and the intra-assay
coefficients of variation were 5.6% (n=4) and 7.7%
(n=4) for sera with testosterone concentrations of 8.71
nmol/L and 4.44 nmol/L, respectively. Testosterone
Can Vet J Volume 35, March 1994

Figure 2. Section of testicular tissue from an 60,XXY,t(1 ;29)
Charolais bull stained with hematoxylin-periodic acid-Schiff.
Seminiferous tubules are partially collapsed, contain few
Sertoli cells, and are devoid of germinal epithelium. The
interstitial tissue contains normal appearing Leydig cells,
which are proportionately inordinately abundant compared to
seminiferous tubules. Bar scale depicts 100 l,m.
concentrations measured over the six-hour period ranged
from 4.82 nmol/L to 12.01 nmol/L. There was a good
response to the GnRH stimulation test, with testosterone
concentrations increasing from 12.01 nmol/L at
zero time to 17.88 nmol/L at two hours after injection of
the GnRH.
The day after the blood sampling for the testosterone
analysis and the GnRH stimulation test was completed,
the bull was castrated. Tissue segments were taken in the
field from the dorsal, central, and ventral regions of
each testis and from the caput, corpus, and cauda epididymis.
They were immediately fixed in Bouin's and
Helly's solutions. The fixed tissue segments were paraffin
embedded, sectioned, and stained for histological study
by light microscopy according to routine methods.
The testicles obtained at castration and dissected
free of the vaginal tunics and epididymides had a combined
weight 80.3 g. The combined weight of the epididymides
was 30.3 g. The left testis was 3.5 X 3.2 X
6.0 cm and the right 3.3 X 3.1 X 6.2 cm in size.
Histologically, the testicular tissue was mainly composed
of normal appearing Leydig cells with a minor
component of poorly developed, partially collapsed,
seminiferous tubules. The seminiferous tubules had a
thick wrinkled basement membrane and contained only
a few Sertoli cells. No germinal cells could be found in
any of the seminiferous tubules (Figure 2). Histologically,
cauda epididymal tissue had a normal appearance; however,
the lumen was completely devoid of spermatozoa.
Can Vet J Volume 35, February 1994
In comparison to his male herd mates and to male
members of his breed in general, the bull appeared
relatively short in height and length. However, no objective
data were obtained on the bull's length, and a shoulder
height of 142 cm is at the low end of normal range.
The bull's masculine facial appearance, general muscling,
and heavy muscling of the dorsal neck region was in contrast
to the feminine appearance of a 61,XXY bull seen
by Dunn et al (2).
The small scrotal circumference measurement is
likely attributable to a decrease in seminiferous tubule
length and diameter, as reported for a 61,XXY bull by
Logue et al (3). The weights of the testicles and epididymides
were comparable to those reported by Dunn
et al (2). The extremely small testicle size was most
likely due to a partial lack of development of seminiferous
tubules and a total lack of development of germinal
epithelium within the seminiferous tubules. The complete
absence of spermatozoa in a seminal fluid sample confirms
the total lack of germinal tissue in this bull's testicles
and is in agreement with Logue et al (3) and
Dunn et al (2) who reported azoospermia in a 61,XXY
Friesian and a 61,XXY Hereford, respectively. The relative
abundance of normal appearing Leydig cells in
comparison to that seen in normal testis tissue is similar
to the situation in the bull reported by Logue et al (3).
The normal size of the bull's penis and accessory
sex glands indicates that there were sufficient levels of
circulating testosterone for the development of these male
characteristics. The serum testosterone concentrations
over the six-hour bleeding period (range 4.82 nmol/L to
12.01 nmol/L) were similar to those of 12 normal bulls
(range 1.87 nmol/L to 21.90 nmol/L) bled over an eighthour
period. The results of the GnRH stimulation test
were also similar to those of 12 normal bulls tested in a
similar manner (unpublished observations) and suggest
a normal responsiveness of Leydig cell tissue in the
secretion of testosterone. Although quantitative estimates
of Leydig cell tissue were not done in this case,
Logue et al (3) reported that Leydig cell volume in a
61,XXY bull was in the normal range.
The karyotype of the bull's dam was 61,XXX and
that of his sire 59,XY,t(1;29). Therefore, we presume that
he inherited the extra X chromosome maternally and the
translocation paternally. Trisomy X cattle have previously
been reported (9-13) as exhibiting limited fertility or,
more commonly, sterility. This dam was obviously fertile
since she produced this calf. She remained on her
home ranch until recently and subsequently produced a
bull calf in both of 1991 and 1992. In January 1993, she
was observed to be in heat after several months without
heats, when, presumably, she was pregnant. By contrast,
Pinheiro et al (14) described a heifer with a 60,XXX,t(1;29)
karyotype that appeared to be infertile, since she was
never observed in heat.
Klinefelter's syndrome appears to be a very rare occurrence.
The few cases (1-3) discussed here were all
cited as rare occurrences, found by chance, and in a chromosome
study of young bulls, including 51 with smaller
than average testicles, we found no cases (15). However,
we recently found a case of 61,XXY in a Simmental calf
during screening of a herd for the frequency of 14;20
translocation (unpublished data). The previously reported
cases (1-3) and the Simmental appear to be the result of
183

nondisjunction during gametogenesis, whereas the
Charolais described in this paper inherited the extra
X chromosome from his dam. His clinical phenotype
appears to be no different from those described
previously, however, even though he also carries a 1;29
translocation.
Acknowledgments
We thank Susan Cook for conducting the testosterone
assays and Bill Kowalenko for supervising weighing and
care of the bull at the Saskatoon R.O.P. Test Station. We
also thank the veterinarians and staff at Intercontinental
Packers for assistance in collecting samples at slaughter.
cvI
References
1. Finger KH, Herzog A, Hohn H, Rieck GW. Fortschritte auf dem
Gebiet der Zytogenetik (Chromosomenpathologie) in der
Veterinarmedizin. Giessner Beitr Erbpath Zuchthyg 1991;
2/3: 13-30.
2. Dunn HO, Lein DH, McEntee K. Testicular hypoplasia in a
Hereford bull with 61,XXY karyotype: the bovine counterpart of
human Klinefelter's syndrome. Cornell Vet 1980; 70: 137-146.
3. Logue DN, Harvey MJA, Munro CD, Lennox B. Hormonal and histological
studies in a 6lXXY bull. Vet Rec 1979; 104: 500-503.
4. Schmutz SM, Moker JS. Impact of a 1;29 Robertsonian translocation
on a herd of purebred beef cattle. Can J Anim Sci 1989;
69: 891-896.
5. Coulter GH, Mapletoft RJ, Kozub GC, Cates WF. Scrotal circumference
of two-year-old bulls of several beef breeds. Theriogenology
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6. Coates JW, Schmutz SM, Rousseaux CG. A survey of malformed
aborted bovine fetuses, stillbirths and nonviable neonates for
abnormal karyotypes. Can J Vet Res 1988; 52: 258-263.
7. Seabright M. A rapid banding technique for human chromosomes.
Lancet 1971; 2: 971-972.
8. Cook SJ, Rawlings NC, Kennedy RI. Quantitation of six androgens
by combined high performance liquid chromatography and
radioimmunoassay. Steroids 1982; 40: 369-380.
9. Rieck GW, Hohn H, Herzog A. X-trisomie beim Rind mit
Anzeichen familiarer Disposition fur Meiosestorungen.
Cytogenetics 1970; 9: 401-409.
10. Norberg HS, Refsdal AO, Garm ON, Nes N. A case report on
X-trisomy in cattle. Hereditas 1976; 82: 69-72.
11. Buoen LC, Seguin BE, Weber AF, Shoffner RN. X-trisomy
karyotype and associated infertility in a Holstein heifer. J Am Vet
Med Assoc 1981; 179: 808-811.
12. Swartz HA, Vogt DW. Chromosome abnormalities as a cause
of reproductive inefficiency in heifers. J Hered 1983; 74: 320-324.
13. King WA, Linares T. A cytogenetic study of repeat-breeder
heifers and their embryos. Can Vet J 1983; 24: 112-115.
14. Pinheiro LEL, Almeida IL Jr, Garcia JM, Basrur PK. Trisomy X
and 1/29 translocation in infertile heifers. Theriogenology 1987;
28: 891-898.
15. Schmutz SM, Flood P, Moker J, Barth A, Mapletoft R, Cates W.
Incidence of chromosomal anomalies among western Canadian beef
cattle. Can J Anim Sci 1990; 70: 779-783.
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184 Can Vet J Volume 35, February 1994