107

J Anat. Soc. India 50(2) 107-111 (2001)

Deterioration Of Sperm Morphology In Men Exposed To High

Temperature
Dada, R.; *Gupta, N.P., Kucheria, K.

Department of Anatomy & *Urology, All India Institute of Medical Sciences, New Delhi-110029, INDIA.

Abstract. Occupational exposure to high temperatures adversely affects testicular function causing partial or complete
spermatogenic arrest. This leads to oligoasthenoteratozoospermia (OAT) and azoospermia. This study reiterates that exposure to high
temperture causes deterioration in sperm morphology and impairs motility. This inverse relationship of sperm function with elevated
temperature has implication in clinical medicine both in understanding pathological states and for therapeutic measures.

Key words : Azoospermia, Oligozoospermia, Oligoasthenoteratozoospermia, Hyperthermia

Introduction : straight and regular in outline. It is aligned with the

About ten percent of males in the reproductive
age group have severe defect in sperm production.
Sperms are produced by a highly complex process
of spermatogenesis. Any partial or complete
disruption of this process results in spermatogenic
arrest and finally leads to oligozoospermia and
azoospermia respectively. In oligozoospermia the
sperm concentration is less than 200 million/ml and
in azoospermia there is complete absence of
sperms in the semen.
For normal fertility, a man requires normal
spermatogenesis, successful epididymal storage
and normal sperm transport. Based on these
functions the causes of infertility can be secretory or
excretory. Secretory causes are due to
spermatogenic arrest while in excretory cause,
spermatogenesis is normal but there is obstruction
to the outflow of spermatozoa. Besides this the
fertility in males depends on normal linear
progressive sperm motility and normal morphology.
Sperm motility is the best predictor of fertility
potential in man.
Several conditions can affect sperm motility
and production of normal sperms. Some of the
known conditions are exposure to high temperature,
deletions of azoospermia factor loci (AZFd) and
iatrogenic causes.
The normal human sperm measures 50-60 mm
in length and has head, neck, middle piece and tail.
The head is oval in shape and is 3-5 mm in length
and 2-3 mm in width. The midpiece is slender,
long axis of the head, and is approximately 7-8 mm
in length. Its width is about one third of the head.
The tail is slender, straight and regular in outline and
is 40-45 mm in length.
In normal conditions testicular temperture is
maintained 3°C lower than the core body
temperature and is an important prerequisite for
efficient spermatogenesis. Germ cells and Sertoli
cells are highly sensitive to elevated temperature.
Various occupational factors and
environmental agents have been shown to have
deleterious effect on male reproductive function and
it is difficult to elucidate the role of a single agent.
Several confounding factors related to diet, life
style, stress and socioeconomic status also affect
semen quality.
Though reports are available regarding the
adverse effects of hyperthermia on semen quality
but very few of these highlight the effect of high
temperature on sperm morphology. The present
paper reports the presence of increased number of
morphologically abnormal sperms with impaired
motility in males with occupational exposure to high
temperature.
Materials and Methods :
Ninety two males within the age group of 22-35
years who were diagnosed as infertile at Urology
clinic of All India Institute of Medical Sciences, New
Delhi were included for this study. Couples are said
to be infertile if the wife is unable to conceive after 1

J. Anat. Soc. India 50(2) 107-111 (2001)

108
year of regular unprotected intercourse. Twenty five
fertile men whose wives had conceived within 1 yer
of marriage were controls (control group).
Information was recorded from patients and controls
about their occupation, type of work performed,
period of cohabitation, and recent illness,
medication taken, exposure to irradiation,
consumption of alcohol, smoking, eating habits, use
of recreational drugs and family history in a
predesigned performa.
Detailed general physical examination was
performed for each patient. FSH and LH levels were
assessed for ech patient to determine the testicular
damage and to confirm that the cause of infertility
was secretory.
Semen samples were collected in a room near
the laboratory and analysed for each patient. A
minimum of 2 semen samples were collected at 1
month interval. For analyses, semen samples were
allowed to liquefy at room temperature and seminal
volume, pH, sperm count, motility and morphology
were analysed according to guidelines in WHO
manual (1992).
For assessing the sperm motility the
microscope field was examined systematically and
motility of each spermatozoa encountered was
classified as A, B, C or D. Rapid linear progressive
motility was motility grade A, while sperms with slow
or sluggish linear or non linear motility were
classified as having motility grade B. Non
progressive motility was grade C and immotile
sperms were classified as grade D.
For studying the sperm morphology a drop of
semen was smeared on a clean glass slide. The
smear was then air dried and fixed in a mixture of
equal parts of ethanol and ether. The slides were
then stained with 2% Giemsa for 30 minutes. Dried
stained slides were scanned under oil immersion
x100 objective to assess for morphological
abnormalities of sperm head, midpiece and tail. A
total of 100 sperms per sample were classified
according to their morphology such as small, pin,
large amorphous, tapered, double and triple head,
dilated or bent midpiece and coiled or short thick
tail.
The mean and standard deviation were
High Temperature & Sperm Morphology
claculated using standard statistical method and
t test to study the significance from two samples
assuming unequal variances.
Results :
Semen analysis was done in 92 infertile males
and 25 controls. Mean age of the patients was 32
years. Of the 92 infertile males, 80 males had no
sperms in their ejaculate and were classified as
azoospermic. Twelve males had a total sperm count
less than 20 million sperms and were classified as
oligozoospermic. In two of the twelve
oligozoospermic males the total mean sperm count
was 15 million with 75% of the sperms showing
normal morphology. The mean seminal volume was
4±1.8 ml and the mean pH was 7.2 ± 0.08. These
sperms had oval head with acrosomal cap covering
more than one third of the head surface. The
midpiece was cylinderical in shape and its width was
less than one third the width of the head. The tails
were long, slender and straight. About 20% sperms
showed abnormal morphology.
Remaining 10 oligozoospermic males were
found to have a very high percentage of
morphologically abnormal sperms with impaired
motility a picture characteristic of OAT and formed
OAT group. These OAT cases were analyzed in
detail and results were compared with control group
(Table 1). Semen volume and pH in OAT group were
slightly lower than the control group, which was not
significant (p>0.05). The average total sperm count
in OAT group was 12 million which was significantly
(p<0.05) lower than the control group. The mean
percentage of sperms with morphological
abnormalities (figure 1 and 2) such as thick coiled
tail, amorphous head, tapered head, double head
and dilated midpiece were significantly higher in
OAT group as compared to the control group (p <
0.05). The mean percentage of sperms with motility
grade C and D were significantly higher in OAT
group than in the control group (p < 0.05) and
motility grade A was significantly lower than the
control group (p < 0.05). The mean percentage of
sperms with motility grade B was lower than the
control group which was not significant (p > 0.05).
The mean FSH and LH levels (21.2 ± 2 mlu/ml
and 13.6±1 mIu/ml) were significantly higher
J. Anat. Soc. India 50(2) 107-111 (2001)
Dada, R. et al 109

TABLE 1
Parameters OAT Group Control Group t-Test
Mean SD Mean SD
Volume (ml) 3 1.2 3.2 1 NS
pH 7.21 0.08 7.24 0.09 NS
Total Sperm Count (million) 12 3.4 63 22.6 S
Abnormal morphology (%) 86.2 2.24 18.8 1.67 S
Thick coiled tail (%) 37.6 7.7 3.45 1.44 S
Amorphous head (%) 25.6 5.18 5.4 0.79 S
Tapered head (%) 23 5.83 6.15 0.07 S
Pinpoint head (%) 6.14 1.14 1.75 0.39 S
Dilated midpiece (%) 4.61 0.45 1.65 0.26 S
Short thick tail (%) 3 1.04 1.4 0.05 NS
Motility grade D (%) 44 7.23 1.9 0.58 S
Motility grade C (%) 39 5.09 4.4 0.65 S
Motility grade B (%) 15 3.16 20.95 1.95 NS
Motility grade A (%) 2 0.94 72.7 2.26 S
NS-Not Significant S-Significant

(p<0.05) in the OAT group males as compared to (DBCP) exposure (n=1) and congenital causes such
the control group (3.54 ± 0.29) mlu/ml and 6.4 ± 0.1 as undescended testes (n=5), hypogonadism (n=14)
mlu/ml). and idiopathic (n=17).

Correlation With Aetiological Factors Discussion :

In 11 azoospermic and 9 males with OAT, there
was one common underlying aetiological factor.
These males had been exposed to very high
temperature at their work place for a period of 5-15
years. They were working in blast furnace (n=6),
Cement, brick, glass and plywood preparation
factory (n=6), melting tar (n=5), Dyers (n=3).
Occupational exposure to consistently high
environmental temperatures led to increased
intratesticular temperatures in these men. There
was no significant history of smoking, medication,
viral illness or use of any recreational drugs in these
males. In the rest of the 69 azoospermic males and
2 oligozoospermic males there were other
aetiological factors which might have led to
complete or partial spermatogenic arrest and
resulted in azoospermia and oligozoospermia. The
causes for spermatogenic arrest in these men were
infections such as tuberculosis (n=5), mumps (n=5),
diabetes mellitus (n=2), iatrogenic (n=3), genetic
factors (n=18), trauma (n=1), dibromochloropropane
J. Anat. Soc. India 50(2) 107-111 (2001)
Temperature of the scrotum is 3°C lower than
the core body temperature and this is an important
prerequisite for optimal spermatogenesis. The germ
cells and sertoli cells are highly sensitive to elevated
temperature which causes partial or complete
spermatogenic arrest (Martin et al, 1993). Previous
studies have shown that some men have intrinsic
defect in scrotal thermoregulation and about 80% of
men with oligozoospermia have elevated scrotal
temperatures (Zorgniotti and Sealfon, 1988). Even in
normal fertile males fever, high summer
temperatures and frequent hot baths, saunas are
known to result in destruction of germinal epithelium
and also induces transient oligozoospermia
(Bedford, 1991; Miusset and Bujan, 1994). Bedford
(1991) proposed that cauda epididymis is sensitive
to high temperature. High scrotal temperature
causes rapid disruption of absorptive and secretory
function of cauda epithelium thereby changing
protein composition of cauda fluid and causing
reduction of its storage capacity. In present study 20
110
infertile males had exposure to high temperature at
their work place for a mean period of 8.5 years. It is
possible that in these men exposure of testes to
high environmental temperature led to an elevation
of intratesticular temperature. This elevation of
intratesticular temperature might have impaired
spermatogenesis and led to production of
morphologically abnormal sperms with impaired
motility to complete absence of sperms in the
semen. Dyers, welders, steel and furnance workers
are exposed to high temperatures at their workplace
and are reported to have impaired spermatogenesis
(Tas et al, 1996).
Available literature suggests that short term
exposure to high temperature may cause reversible
changes in the seminiferous tubules whereas
chronic exposure for 5-7 years had OAT while those
exposed for 12-15 years had azoospermia.
Wang et al. (1997) reported that elevation of
testicular temperature 1°C above the baseline
depressess spermatogenesis by 14% and thus
decreases sperm output. Exposure to high
temperature also results in modification of sperm
morphology. It is characterized by increase in the
number of morphologically abnormal sperms. The
mean value of sperms with abnormal morphology
rises from 30 to 60% within 6-8 months of exposure
to high temperature (Wang et al, 1997). They
postulated that heating the testes induced a
depression not only in the amount but also in the
quality of sperm output.
Though a number of studies have reported
about deterioration in semen quality on exposure to
high temperature but to the best of our knowledge
very few studies have reported about the increase in
specific morphological abnormalities of sperms. In
our study we found that 86.2% sperms had
abnormal morphology. The most predominant
abnormality was thick coiled tail (37.6%) and
amorphous head (25.6%). Though percentage of
sperms with morphological abnormalities rises
sharply within 6-8 months of exposure (Wang et al,
1997; Mieusset et al, 1991; Levine et al, 1992), in
the present study the cases had exposure for the
mean period of 8.5 years. Wang et al. (1997)
explained that elevated testiculoepididymal
temperature decreases the synthesis of sperm
High Temperature & Sperm Morphology
membrane coating protein which in turn results in
the production of morphologically abnormal sperms.
Only 2% sperms showed linear progressive
motility and whereas majority (83%) of sperms had
impaired motility (83%) of sperms had impaired
motility (grade C and D). Increase in number of
morphologically abnormal sperms results in
impaired motility as normal intact sperm
morphology is prerequisite for linear progressive
motility. Gandini et al. (2000) postulated that sperm
function is strictly correlated with sperm morphology
and that sperm motility is the best predictor of
fertility potential in man.
Functional and fully differentiated Sertoli cells
are critical for development of quantitatively and
qualitatively normal spermatogenesis. They also
provide structural and functional support to the
developing and differentiating germ cells as each
Sertoli cell is in contact with 47 germ cells and 5
other Sertoli cells (Orth et al, 1988). High testicular
temperature damages the Sertoli cells and their
number decreases impairing spermatogenesis. This
leads to a decrease in the number of germ cells with
incomplete differentiation (Steger at al, 1999).
Damaged Sertoli cells produce decreased amount of
inhibin which decreases negative feedback on
pituitary leading to inrease in FSH level. This
increase in FSH level is directly proportional to the
Sertoli cell damage that indicates the severity of
testicular damage. FSH levels are the most
important endocrine parameter to evaluate testicular
function (Bergmann et al, 1999). FSH and LH levels
were elevated in azoospermia and in OAT cases.
The difference in FSH and LH levels between OAT
group and control group were statistically significant
(p < 0.05).
One of the well known mechanism to explain
spermatogenic impairment due to hyperthermia is
activation of p53, a tumour suppressor gene which
is expressed in testes (Rogel et al, 1985, Almon et
al, 1993). Its level of expression is highest in
pachytene spermatocytes (Schwartz et al, 1993).
High scrotal temperatures cause condensation of
nuclear chromatin which causes p53 activation
which leads to cell cycle arrest. This prevents clonal
proliferation of germ cells with damaged DNA.
Morgentaler et al. (1999) postulated that p53 might
J. Anat. Soc. India 50(2) 107-111 (2001)
Dada, R. et al
play a role in heat induced germ cell apoptosis. p53
is located on the nuclear membrane in the normal
germ cells and is involved in quality control of germ
cells. It translocates to the nucleoplasm with heat
induced nuclear damage and induces germ cell
apoptosis (Yin et at, 1997).
In the past decade, male reproductive health
had declined with marked increase in the population
of subfertile males (Carlsen et al, 1992). The sperm
count has been falling at an alarming rate of 2% per
annum for the last 20 years. This is believed to be
due to increase in global temperature and
environmental pollution. There has been an yearly
decline of 2.6%, 0.3% and 0.7% in sperm
concentration, sperms with normal motility and
sperms with normal morphology (Tas et al, 1996).
Thus, the testes is remarkable as a biological
system for its functional regulation by temperature.
The testes function optimally at relatively cool
temperature and high testicular temperatures impair
spermatogenesis leading to OAT and azoospermia
as has been seen in the present study. Observed
temperature sensitivity of testes has implications in
clinical medicine both in understanding pathological
state and for therapeutic measures. Thus exposure
to high temperatures at the workplace is an
occupational hazard and leads to deterioration of
sperm morphology and impaired motility which may
result in infertility.
Acknowledgements :
One of the authors R Dada is receiving
fellowship from Council of Scientific and Industrial
Research (CSIR) and wishes to thank CSIR for its
financial support.
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Opp. 110 High Temperature & Sperm Morphology

Fig. 1. Semen microphotograph from an OAT case. A very high percentage of sperms with
morphological abnormalities (x400).

Fig. 2. Semen microphotograph from OAT case showing A Spermatozoa with coiled tail B Spermatozoa with dilated bent
midpiece C Spermatozoa with double head D Spermatozoa with triple head