THE MALE GENERATIVE CYCLE IN THE

HEDGEHOG; WITH EXPERIMENTS ON THE

FUNCTIONAL CORRELATION BETWEEN THE
ESSENTIAL AND ACCESSORY SEXUAL ORGANS.
BY F. H. A. MARSHALL.

(From the Physiological Laboratory and the School of

Agriculture, Cambridge.)

THE changes which occur during the cestrous or female generative cycle
and the functional correlation subsisting between the various organs
concerned have formed the subject of recent research by several in-
vestigators'. It has been shown that in mammals the processes of
maturation and ovulation are confined normally to certain definitely
restricted periods which usually recur rhythmically, and further, that
the changes which take place in the uterus bear a close relation to
those going on synchronously in the ovaries, this relation being
apparently dependent upon the existence and functional activity of
ovarian internal secretions which circulate in the blood. In the present
paper it is proposed to deal with the corresponding phenomena in the
male organism, and more particularly to describe a series of observations
and experiments upon the reproductive organs of the male hedgehog,
in which the cyclical changes are considerably more pronounced than
they are in most otber animals.

The male generative cycle.

The males of most species of Mammalia do not exhibit the same
degree of sexual periodicity as that shown by the females. In some,
however, there is a definite period of rut at the same time as the sexual
1 Heape. Quart. Journ. Micr. Sci. xrxv. 1900. For further references see Marshall,
The Physiology of Re,production, London, 1910.
248 PHYSIOLOGY OF SEXUAL ORGANS.
season in the female. In such animals mature spermatozoa are pro-
duced at rut and not ordinarily at other times, at least so far as is
known. Probably in the majority of wild animals the reproductive
functions are restricted to recurrent seasons of rut, but in the domestic
animals the tendency towards this limitation is more or less obscured
though still often marked. Thus, in winter or tinder adverse conditions
of environment the sexual capacity may be reduced. Nevertheless
it is undeniable that the power to reproduice is more extended and
less limited by seasonal periodicity among the males of domesticated
animals than it is among related wild varieties or species, and this
increased capacity is probably due to the favourable surroundings and
conditions of comparative luxury amid which domesticated animals ale
reared. The power to generate in the male mammal is apparently
extended equally over the entire rutting season and is not restricted to
recurrent periods comparable to the cestrous periods of the female,
these latter being separated by resting intervals of short duration
though falling within the limits of a single breeding season. It would
seem, however, that there inay be a slight tendency towards the
rhythmical recurrence, at short intervals, of periods of increased sexual
activity in man. In the lower animals, as far as is known, there are no
such periods, the capacitv to breed at a given time (luring rut appearing
to depend upon other factors, and largely upon the extent to which sexual
intercourse has taken place previously duiring the season in question.
The approach of rut is marked by the growth of the generative
organs, and especially of the testes, associated with the formation of
spermatozoa. In some Mammalia these processes are accompanied by
changes in the anatomical position of the testes.
In birds the periodic increase in size undergone by the testes is
always considerable and sometimes enormous. This was pointed out
by Aristotlel. Gadow2 remarks that the testes of the house sparrow,
which during the non-breeding season are often no bigger than mustard-
seeds, at the approach of the pairing time (in April) become as large as
small cherries. Disselhorsts states that in the finch these glands
increase three-hundred-fold. According to Wilson4 the first sign of an
enlargement of the testes takes place about the third week in February
in the red grouse (in Scotland). In May they have increased about
1 Aristotle. De Generatione Animalium. Platt's Translation, Oxford, 1910.
2 Gad ow. Article inNewton's Dictionary of Birds, London, 1896.
3 Disselhorst. Anat. Anz. xxxii. 1908.
4 Wilson. The Grouse in Health and Disease, London, 1911.
 F. H. A. MARSHALL. 249
twenty or thirty times, having in the meantime become white and fatty
in appearance, instead of small and deeply pigmented. I have noted
similar facts in regard to starlings, thrushes, blackbirds, rooks, and
many other common birds. Patten, who has also paid some attention
to this subject states that the maximum size in the testes in many
common birds is reached about a fortnight before the hens first begin
to lay. The increase is partly due to the great enlargement of the
tubules of the gland, this enlargement being correlated with active
sperm proliferation. After the breeding season is over the testes
again become reduced.
In Monotremata, there is said to be a single annual breeding season,
but as there are no vesicule seminales or prostate glands the changes
which occur in the male generative organs must relate almost solely to
the testes. These organs, however, remain permanently in the body
cavity as in lower vertebrates and do not descend into a scrotum during
rut.
Nothing appears to be definitely known about the male generative
cycle in Marsupialia. The testes in this order of mammals remain in
the scrotum throughout the year. Probably however there is a rutting
season corresponding to the female sexual season.
In the Rodentia the testes undergo periodic enlargement in size
and change of position (excepting in the Leporidae in which these
organs appear usually to remain in the scrotum throughout the year).
After rut the testes pass back from the scrotum into the abdominal
cavity where they remain till the onset of the next rutting season.
In most species this begins in the early spring and extends until the
stummer or sometimes later. At any timne during the rutting season
the males are capable of copulation whereas the females experience
rectirrent cestrus interrupted by periods of rest or by pregnancy. During
the height of winter the reproductive capacity of male rodents is
minimal or non-existent, so that even in tame rats or rabbits kept in
captivity it is often hard to find fully developed spermatozoa in the
testes or other parts of the generative tract. Owen2 remarks that
the periodic enlargement of the testes is especially marked in rats and
mice.
In most male Ungulata there is a rutting season occurring syn-
chronously with the female sexual season. In the domesticated kinds,
however, it is evident that in the case of the male the restriction is very
1 Patten. Brit. As8oc. Report, Sheffield Meeting, 1910.
2 Owen. Anatomy of Vertebrata, i. London, 1866.
 250 PHYSIOLOGY OF SEX UAL ORGANS.
indefinite and may be non-existent. This is indicated by the fact that
ewes which abort at mid-pregnancy or even later may come in use
subsequently at an exceptional time and succeed in getting served by
a ram'. There is no doubt however that a ram which has served
the usual number of ewes (abotut forty or fifty) at "tupping time" is
generally exhausted, and that the testes seem to become smaller and
tend to be drawn upward in the scrotum but without passing into the
abdomen. In the boar in a state of domestication there appears to be
no restricted time for rutting, and a similar remark may be made about
the male goat, the bull, and the stallion. On the other hand in all
these animals the sexual capacity is variable, but there is no obvious
periodicity such as occurs in the female animals.
In the wild species the males experience a periodic rut, but the
testes though increasing in size do not appear to change ln position, at
any rate not to any appreciable extent. In some species (e.g. deer) the
activity of the generative organs is accompanied by the appearance of
secondary sexual characters which develop at the approach of the
breeding time and vanish after rut is over. The periodic growth of
the antlers in stags is a familiar instance of this kind of correlation.
The prong-buck (which alone among hollow-horned ruminants usually
sheds its horns) is another example. The accessory sexual glands (e.g.
the perineal glands) in musk deer and other kinds of deer are commonly
believed to serve as a means of attraction between the sexes and are
especially active during rut.
The temporal gland of the male elephant is stated to show an
increased activity during rut, but the periodicity in elephants in
captivity is very irregular and in the great miajority of cases ,they refuse
to breed at all2.
In all Ungulata which experience rut, this season is one of great
sexual excitement. At other times the males of many species herd
together apart from the females.
It is interesting to note that the male camels in the Zoological
Society's Gardens in London experience rut in early spring (or at the
same time as the wild camels in Mongolia) in spite of the differences in
climate and the fact that they are kept in captivity3.
Male Carnivora kept in captivity or under domestication do not
usually experience any definite rut. The dog and cat are apparently
able to breed at all times, but Cocks states that the male wild cat in
1 Marshall. Phil. Trans. B, cxcvi. 1903.
2 Morgan. Experimental Zoology, New York, 1907. 3 Heape. Loc. cit.
 F. H. A. MARSHALL. 251
captivity has a recurrent rutting time during which it calls loudly and
displays great sexual excitement1. Little or nothing is known about
the breeding habits of land Carnivora in their natural state. The males
of seals have a regular rutting time, which in some species at any rate
is a period of complete fasting. Thus it is stated that the male fur
seals after coming to land may live for over a hundred days without
taking food, and that during this period they are constantly engaged
in struggles with other males, finally leaving the shore in a state of
extreme emaciation.
In the Insectivora the periodic changes in the male generative organs
are especially pronounced. Thus, in the mole in December the testes
are on each side of the bladder, and externally it is a matter of great
difficulty to distinguish the males from the females. From January
onwards the mnale generative organs undergo a progressive increase
which culminates in March, when the testes are protruded into sacs
near the base of the tail. The prostate glands also undergo an enormous
enlargement and conceal the bladder at the end of March. Pairing
takes place at about this time, or in April, or sometimes as late as the
first part of May. Subsequently the generative organs become quiescent
and correspondingly reduced, and so the annual cycle is repeated. In
the Cape mole there are probably comparable changes, the convoluted
course of the vas deferens indicating an alteration in the position of the
testes2. In the hedgehog the male organs undergo similar periodic
changes which are described below in dealing with their histology.
The breeding season extends from April until August or September.
The shrew also has a rutting season which may last from April until
November. The testes in this animal become enlarged and pass into
temporary receptacles in the perinaeum, but there is no true scrotum2.
In European genera of Cheiroptera, which hilernate during the
winter, copulation takes place regularly in autumn before they enter on
their winter sleep, the spermatozoa remaining stored up in the uterus
of the female until the succeeding spring. In these animals, therefore,
there is presumably a definite male rutting season.
Among the Primates it has been shown that there is probably
a breeding season for certain species of monkeys in parts of India. It
would seem likely therefore that the males of these species have a
periodicity in rutting3.
1 Cocks. Lettertotheauthor.
2Owen. Loc. cit.
3 Heape. Phil. Trans. B, CLXXXV. 1894; and CLXXXVIII. 1897.
PH. XZLIII. 17
252 PHYSIOLOGY OF SEXUAL ORGANS.
The evidence of a pairing season in primitive man has been
discussed elsewhere. Apart from this question Havelock Ellis and
Percy Costel state that there are indications of a rhythmic reguilarity
in the sexual functions of men under conditions of civilization.

The cycle in the male hedgehog.

That the accessory sexual glands of the male hedgehog undergo
a periodic metamorphosis was briefly commented on by Owen2. More
recently Griffith s3 has described the histological appearances presented
by the hedgehog's prostate glands (luring quiescence and activity, as
well as those of an intermediate stage. Griffiths however does not
deal with the other sexual organs.
In the present investigation hedgehogs were killed at all seasons of
the year, fourteen males showing different degrees of generative activity
having been employed in addition to the animals referred to in the
experimental enquiry described below. The animals killed during
winter were obtained before hibernation and were kept for several
months in captivity. The sexual organs were in nearly all cases
preserved entire (excepting for the copulatory organ) and sections were
afterwards cut.
In this country, according to Millais4, the hedgehog produces
litters in May or June, and again in August or September, but the
condition of the male organs as late as the beginning of October would
seem to indicate the occasional possibility of breeding almost up to
that time. During September or October the sexual organs rapidly
become smaller, until in mid-winter the accessory glands are not easily
discernible. The increase in size begins about the end of March
(shortly after the close of the hibernation period). By the middle of
April it is considerable, the organs having at that time reached about
one-third of their full dimensions. Complete development is attained
to in May; and throughout the remainder of the spring and summer
their condition is approxinmately the same. As remarked by Oudemans5
the sexual organs of the rutting hedgehog in proportion to its size are
more strongly developed than those of any other animal.
1 E ll i s. Psychology of Sex, II. Philadelphia, 1900.
2 Owen. Loc. cit.
3 Griffiths. Journ. of Anat. and Phys. xxiv. 1890.
4 Milla is. The Mammals of Great Britain and Ireland, I. London, 1904.
50 udem a ns. Die Accessorischen GeschlechtsdrWsen der Saugethiere, Haarlem, 189
 F. H. A. MARSHALL. 253
Apart from the testes, the organs concerned with reproduction are
the vesiculae seminales, the prostates, Cowper's glands (in the muscle
of the urethra), and a pair of additional glands (outside the pelvis on
the buttock) considered by Linton, to be sui generis but sometimes
regarded as representing Cowper's glands.
The testes become considerably enlarged with the approach of rut
but apparently not to the same extent as in the mole with which,
according to Regaud2 and Lecaillons, they increase sixty-four times in
size. During the winter the spermatogenetic function is in abeyance,
but active spermatogenesis is going on in April, at the end of which
month numerous free living spermatozoa may be found in the genital
passages. The increase in the dimensions of the testes is due partly to
the growth (both in size and number) of the seminiferous tubules
associated with spermatogenetic tissue, but even more largely to the
proliferation of the interstitial cells. The result of this process is that
during rut the seminiferous tubules are often widely separated, especially
in the more central portion of the testes. Blood vessels of considerable
size are not uncommon in the inter-seminiferous tissue, but they
disappear when retrogression sets in. After October the seminiferous
tubules are brought into almost close contact, and the entire organ
remains comparatively small and compact throughout the remainder of
the period of rest.
The testes do not descend into a scrotum during the rutting season
but occupy sac-like continuations of the abdominal cavity in the
neighbourhood of the perinaeum where during rut they may be detected
from the exterior.
The increase in size of the vesiculae seminales is more noticeable
than that occurring in any of the other sexual organs, because on
dissecting from the ventral surface these glands are seen to stretch so
far forwards as to obscure the neighbouring viscera in the body cavity.
In winter they are hidden by the urinary bladder. The weight of the
vesiculae with their contained secretion from a hedgehog killed in
August was found to be about 6 grams, whereas that of one killed in
January was only 1 gram. Comparisons with other hedgehogs killed
respectively in summer and winter showed that these two animals were
typical in regard to the development of the sexual glands at these
seasons. Near the end of April the vesicula were found to be about
1L i n t o n. Anat. Anz. xxxi. 1907.
2Regaud. C. R. de l'As8oc. Anat. 1904.
3 L ecaillon. C. R. de la Soc. de Biol. LXVI. 1909.
17-2
2054 PHYSIOLOGY OF SEX UAL ORGANS.
one-fifth developed and already to contain the characteristic secretion.
In May they had reached their full size which they retained until
September or October. They contained a secretion during the whole
of that time. After the middle of October (or sometimes a much
earlier date), the glands began rapidly to diminish and the secretion to
become much reduced in quantity.
During the active stage the epithelial cells of the acini were long
and cylindrical, and their protoplasm appeared to contain granules.
At the resting time these cells were smaller and almost cubical in
appearance, as described by Rauther to be their condition in September.
There has been some controversy regarding the function of the
seminal vesicles in mammals. According to one view they are re-
ceptacles for the spermatozoa preparatory to seminal ejaculation. Hence
their name. Thus, Rehfisch2 states that fluids injected into the
testicular end of the vas deferens enter the vesiculae before passing out
by the urethra. According to Misuracas in dogs and cats which do
not possess vesiculv the spermatozoa disappear froin the genital passages
a week after castration, if not sooner, whereas in guinea-pigs which have
large seminal vesicles, living sperms may be found three weeks after
the extirpation of the testes. This is regarded as evidence that the
vesiculae act as seminal reservoirs. In rabbits, which also have no
vesiculae, spermatozoa have been found alive in the vas deferens at
least ten days after castration4. Certain of the older writers state that
they found spermnatozoa in the vesiculae seminales of the mole and the
hedgehog during the breeding season, but these observations do not
appear to have been confirmed5. On the other hand Landwehr6,
Camus and Gley', Sobotta8 (for rodents), and Rauther9 (for rodents
and insectivores) have recorded the recurrence of definite secretions
from the seminal vesicles. According to Kolster'° the epithelial cells
of the vesicule in the elk undergo desquamation during secretory

1 In one case the vesiculn seminales were found much reduced and without any
seeretion as early as the end of August.
2 Behfi s ch. Deutsche med Woch. xxii. 1896.
Misuraca. Rivista sper. di Frenatria, xv. 1890.
M
4 Marshal l and Jolly. Phil. Trans. B, cxcviiI. 1905.
5 For references see Marshall. The Physiology of Reproduction, London, 1910.
6 Landwehr. Pftuiger's Arch. xXIII. 1880.
7 Carnus and Gley. C. R. de la Soc. de Biol. iv. 1897.
8 Sobotta. Arch.f. mikr. Anat. XLV. 1895.
9 Rau the r. Jenaische Ztschr. xxxviI. 1903.
I Kolster. Arch. f. mikr. Anat. LX.
1892.
 F. H. A. MARSHALL. 255
activity. Akutsul also has described these cells as undergoing func-
tional changes, being said to contain more plasma during rest than
during activity.
II have made microscopical examinations of the vesiculae seminales
of the hedgehog at all seasons of the year, and in no instance have I
been able to discover spermatozoa, even after the most careful search
and'during the middle of the breeding season. On the other hand I
have never failed to find abundant evidence of secretory activity during
rut (as already mentioned). The glands contained a large quantity of
a white or yellowish-white fluid, which on examination under the
microscope was found to consist mainly of very numerous floating
particles resembling irregularly shaped crystals. They are different in
appearance from Bottscher's crystals which are formed partly from
prostatic spermine, and from the crystals of Lubarsch which occur in
the tubules of the testis. They do not appear to have been recorded as
occurring in the vesiculae of rodents or other mammals, and so are
possibly peculiar to those animals which undergo a very extensive
development of the vesiculae during rut. Dr Hopkins undertook to
exanmine the crystals chemically, and his observations are described in
an Addendum to this paper.
From the above described observations it is clear that in the
hedgehog if not in other mammals the vesiculae senminales are essentially
secretory glands probably contributing to the formation of semen, and
are not receptacles for storing sperinatozoa. Their great size and the
quantity of secretion formed in them seem, however, to suggest that
in the hedgehog at least they must possess some more important
function than that merely of diluting the seminal fluid.
The changes undergone by the prostate have been described by
Griffiths2 with whose observations mine are in general agreement.
During winter the organ appears as a small glandular structure at the
beginning of the' urethral canal. The epithelial cells of the tubules are
cubical in shape. In March the organ is two or three times enlarged,
the tubules of which it is composed being much more numerous and
extensive. Meanwhile the epithelial cells are in process of elongation.
During the active stage the prostate gland is from eight to ten times
its former size (when quiescent). The tubules contain a secretion, and
the cells forming their epithelial lining are columnar. In the autumn
when the breeding season is over the organ undergoes retrogression,
the tubules becoming onice more reduced. The changes which occur
1 A k u t s u. Pflaiger's Arch. xcvi. 1903. 2 Griffiths. Loc. cit.
256 26 PHYSIOLOGY OF SEXUAL ORGANS.
are pronounced, but not so much so as in the case of the vesicule
seminales referred to above.
The second prostates or so-called Cowper's glands (which lie outside
of the pelvis close to the root of the penis) also undergo cyclical
changes as described by Linton'. During the breedinig season the
secretory acini of these glands produce a quantity of fluid containing
small round bodies the nature of which has not been determined.
During the non-breeding period the secretory tubules are inactive and
much reduced in size and the gland consists largely of connective
tissue.
The Cowper's glands of Leydig (which are probably homologous
with Cowper's glands in other mammals) likewise appear to pass
through periodic changes of activity and rest. During the breeding
season there is a distinct increase in the size of these glands.

The correlation between the sexual organs in the male hedgehog.

In view of the exceptionally great changes undergone periodically
by the sexual organs in the male hedgehog it seemed that this animal
was peculiarly suitable for an investigation undertaken to elucidate the
nature and extent of the fiunctional correlation subsisting between these
organs. The following series of experiments in which the animals were
castrated or vasectomised, either completely or unilaterally, was ac-
cordingly carried out:
(1) Both testes were removed from a hedgehog in March prior to
the beginning of the breeding season. At the time of the operation2 the
accessory generative organs had not begun their periodic increase in size,
at least so far as could be observed. Two months later it was found
that these organs were still undeveloped and quiescent, but no changes
that could definitely be said to be atrophic were noticed.
(2) Both testes were removed from a hedgehog in the very early
part of the breeding season, the accessory generative organs being about
half developed. Six weeks later these organs were found to be practi-
cally unaltered as regards size, although those of other, unoperated
hedgehogs were at that time fully developed and functional. It was
clear, therefore, that the extirpation of the testes had arrested the further
growth of the vesiculae seminales and other sexual organs.
Linton. Loc. cit.
2 Since castration in the hedgehog involves an abdominal operation, it was possible to
se something of the accessory organs at the time when the testes were removed.
 F. H. A. MARSHALL. 257
(3) The left testis only was removed from a hedgehog early in the
commencement of the breeding season when the sexual organs had only
lately beaun their periodic growth. Three months afterwards both
vesiculae were found to be equally developed and actively secreting.
The other accessory organs were apparently normal. The remaining
testis (right) was well developed.
(4) The right testis only was removed very early in the breeding
season as in the preceding experiment. The result also was apparently
identical, both vesiculae being found to be normally developed and in a
state of active secretion three months later when the hedgehog was
killed.
(5) This experiment was identical with the preceding one, the
right testis only being removed. The result after three months was
similar.
(6) Both vasa deferentia were transected about the middle of their
length in a male hedgehog in March before the beginning of the breed-
ing season. Four months later the testes and vesiculae seminales
appeared superficially only to differ from the normal in being somewhat
smalL The vesiculee contained the characteristic secretion. Spermato-
genesis appeared to have begun in the testes, but the fate of mature
spermatozoa, if such were produced, was not determined.
(7) The right vas deferens only was cut very early in the
beginning of the breeding season. Three months later the vesicula
seminales were found to be normal and in a condition of active secretion,
the other glands being also normal as far as was observed. The right
testis, however, was found to be only about two-thirds the size of the
left one, this result having perhaps been brought about through the
cessation of sperm production resulting from the. absence of outlet for
the mature spermatozoa.
(8 and 9) In these experiments only one vas deferens was tran-
sected. The vesiculae seminales appeared to continue to undergo
growth, but as the hedgehogs, although completely recovering from the
operations, died from some other cause within a month, the results
cannot be regarded as satisfactory.
In all the vasectomy experiments the spermatic cord and vascular
connections of the testis were left intact.
The experiments demonstrated very clearly the existence of a close
functional correlation between the essential and accessory generative
organs in the male hedgehog. Moreover, since unilateral castration had
no effect in arresting the remarkable periodic growth of the vesiculm
 258 PHYSIOLOGY OF SEXUAL ORGANS.
and other organs, and since vasectomy, either unilateral or double,
likewise had no inhibitory influence, there is strong evidence that the
cyclical changes in the accessory glands are not brought about through
stimuli set up by the external testicular secretions or by means of the
nervous system, but by substances carried from the testes in the
circulating blood.
The results of the experiments, therefore, are in general agreement
with the theory adopted by Shattock and Seligmann', Nussbaum2,
Steinach8, and others that the testis is an organ of internal secretion.
Assuming that this is the case, it is clear that in the hedgehog
the internal testicular secretion iacts periodically and not continuously.
Moreover, it would seem probable that the great development of
interstitial tissue which occurs at the beginning of the breeding season
is in correlation with the contemporaneous hypertrophy of the vesicule
seminales and other accessory glands. If this is so, it is in conformity
with the view adopted on experimental evidence by Ancel and Bouin4
and by Copeman5, that the problematical internal secretion of the
testis is elaborated by the interstitial cells and not by the spermato-
genetic tissue6.
CONCLUSIONS.
The following are the main conclusions which have been reached
(1) The males of most if not all wild mammals experience a definite
sexual cycle, the testes and accessory generative organs passing through
alternate periods of rest and activity, but in the domesticated varieties
and in man the period of rest is liable to be much reduced or may be
absent altogether.
I Shattock and Seligmann. Proc. Roy. Soc. LXXII. 1904.
2 Nussbaum. Ergeb. der Anat. und Entwick. xv. 1905.
3 Steinach. Zntrlb.f. Physiol. 1910, No. 13.
4 Ancel and Bouin. Arch. de Zool. Exp. i. 1903.
6 Copeman. In a Paper read before the Physiol. Soc. 1908 but not published.
6 My experiments shed no further light on the precise functions of the various
accessory generative glands in animals. Walker, however, has recently published the
results of an investigation on this subject. His experiments were upon white rats. The
general conclusions reached are that whereas extirpation of the prostate together with the
vesicule seminales (though not inhibiting sexual capacity) causes sterility, the removal of
either of these organs independently does not necessarily (or even generally) produce
sterility (Johns Hopkins Hosp. Reports, xvi. 1911). Iwan off, however, has shown that
the females of various rodents could be successfully artificially inseminated by the
injection of spermatozoa obtained directly from the testes and therefore unmixed with the
secretions of the accessory glands (Journ. de Phys. et de Path. Gen. ii. 1900).
 F. H. A. MARSHALL. 259
(2) In the male hedgehog the period of activity extends usutally
from about the beginning of April until sometime in September, the
accessory generative organs, and more particularly the vesiculio seminales,
acquiring enormous proportions at this time, after which they become
very much reduced in size until the approach of the next breeding
season.
(3) The vesicul] seminales are secretory glands, and do not contain
spermatozoa either in the breeding or in the non-breeding season.
(4) Complete castration during the period of rest prevents the
seasonal development of the vesiculae seminales and other accessory
male organs, or, if such development has already begun, arrests its
further progress. IUnilateral castration, however, does not inihibit the
growth of the accessory organs and has no effect on the symmetrical
arrangenment of these organs.
(5) Vasectomy, either unilateral or double, does not inhibit the
growth of the vesiculae seminales or other accessory organs, but uni-
lateral vasectomy may prevent the full development of the testis on the
side of the operation, as compared in size with the testis on the
unoperated side.
(6) The periodic development of the accessory organs is not due to
stimuli set up by sperm ejaculation, but is probably the result of an
internal testicular secretion elaborated in the interstitial tissue during
the season of generative activity.
The expenses of the investigation were partly defrayed by a grant from the Government
Grant Committee.

Addendum: A Note on the Chemistry of the Vesicular

Fluid of the Hedgehog.
BY F. GOWLAND HOPKINS.
From the glairy milky fluid expressed from the vesicule seminales,
the suspended crystalloid material observed by Dr Marshall and men-
tioned in his text was found to separate easily on centrifuging, the
supernatant fluid being then clear. By successive treatment with
normal salt solution and water (in neither of which is it soluble) the
centrifuiged precipitate could be entirely freed from the other con-
stituents of the vesicular fluid. Microscopically it was composed of
transparent, somewhat irregular, masses closely resembling small,
'260 PHYSIOLOGY OF SEXUAL ORGANS.
ill-formed, crystals of edestin. On chemical examination it proved to
be a protein containing phosphorus. It exhibited typically the biuret,
xanthoproteic, Millon, and glyoxylic reactions, while the residue ob-
tained after fusing 0-1 grni. with carbonate of soda and nitrate of potash
gave, when dissolved in nitric acid, a well-marked yellow precipitate
with ammonium molybdate. Somewhat remarkable is the fact that
even prolonged boiling with water affected neither the form, nor the
complete transparency, of the crystalloid masses. Their appearance
was also unaltered by prolonged standing under alcohol. The protein
differs from most phospho-proteins in its resistance to solution in
alkalies. In sodium carbonate solutions it scarcely dissolved at all.
In dilute sodium hydrate it could be dissolved, but apparently not
without decomposition. On acidifying its solutions in the latter an
amorphous precipitate fell, and there was well-marked evolution of
hydrogen sulphide.
Its insolubility in alkali carbonates explains its separation from the
vesicular fluid, which itself has a slightly alkaline reaction. On boiling
with dilute hydrochloric acid the crystals slowly dissolved, and the
solution reduced Fehling's reagent.
The material available was not sufficient for a quantitative analysis,
and I was unable to decide whether purin groupings were present.
This crystalloid insoluble substance certainly formed a large pro-
portion of the total proteins of the vesicular fluid, and the separation of
such a protein in quantity from a physiological secretion in situ seems
to be a soimewhat remarkable and exceptional phenomenon. In the
clear portion of the fluid albumens and globulins were present in small
amount.