JF 871 G5 opy 1 ^^^| A Study of Some Factors Influencing Fertility and Sterility in the Bull BY HERBERT L. GILMAN Reprinted from the Annual Report of the New York State Veterinary College 1921-22 A Study of Some Factors Influencing Fertility and Sterility in the Bull A THESIS Presented to the Faculty of the Graduate School of Cornell University for the Degree of Doctor of Philosophy BY HERBERT LESTER GILMAN, D. V. M., M. S. Reprinted from the Annual Report of the New York State Veterinary College ALBANY J. B. LYON COMPANY, PRINTERS 1922 SFsn A STUDY OF SOME FACTORS INFLUENCING FERTILITY AND STERILITY IN THE BULL ^y • — '<, HERBERT L. GILMAX Veterinary Experiment Station, Cornell University Normal reproduction is the fundamental foundation upon which the entire cattle industry rests. For this reason, any factor capable of interfering with it is a detriment to the industry, and a matter of prime importance to the breeder and the veterinarian. With the relative increase in number and value of cattle, and the fact that the profession is depending more and more on this in- dustry for a livelihood, these problems are assuming greater im- portance. The part played by the bull has been emphasized entirely too little, with the result that, as in human medicine, many fail to appreciate the effects of sterility or lowered fertility in the male. The part played by the sire in the spread of genital infections, though discussed frequently, has received little sys- tematic investigation. The bull must be regarded as at least half the herd, not only from the standpoint of the characters he imprints upon his prog- eny, but because of his relation to the reproductive efficiency in the herd. It seems quite probable that he does disseminate during copulation, infection associated with the genital organs, with the result that the bull is a very important factor in a study of the subject. Too frequently, his ability to copulate in an apparently normal manner, is taken as a standard of fertility. Gross changes in his genitalia, or the absence of spermatozoa from the semen are given due consideration, while other more obscure abnor- malities are not looked for nor regarded in their proper light. Neither fertility nor sterility are always absolute, but the terms should be used relatively inasmuch as we may have all degrees of infertility or impotency. All too frequently we forget the many delicate and intricate mechanisms involved in the reproductive process, with the result that many phases of the problem are neg- lected or disregarded. The genital organs work as a unit, each part of which must function in perfect accord with the others to the end that full fertility may result. The physiological factors involved in the formation of the semen are too little understood, or at best, our knowledge regarding them is more or less hazy. The purposes of the present work have been: (1) to summarize the work so far done on the subject. (2) to review briefly the known facts throwing light on the anatomy and physiology of the male genital organs, (3) to carry out systematic studies upon the pathology and bacteriology of the genital tract of the bull, and (4) to ascertain if possible whether the bull is a disseminator of those infections which interfere with reproduction in the female. [3l The work has been carried on for the tnosl pari from the point a Laboratory man cooperating with clinicians. .No attempl is made in this paper to give detailed clinical data, methods for physical examinations, etc. There are included many state- ments ami some data given in a preliminary article on the sub- ject. While the subjed is broad in its scope, in fad too broad for greal detail, it is hoped thai a start has been made toward future and more detailed investigations. Bistort References to, and investigations relating to, the pari played by the hull in the process of reproduction in the herd, and in the spread of genital infections, have been limited Largely to those phenomena caused by Bad. abortum. Bang (] I originally called attention to the possibility of the male transmitting the organism discovered by him. hut he reached no definite conclusion on the subject, dames Law (2) writing on contagious abortion in cows, early suspected this possibility when stating under " casual infec- tious." thai " In a case winch came under the observation of the writer recently, a family cow. kept in a barn where no abortion had previously occurred, was taken for service to a hull in a herd where abortion was prevailing, and though she was only present at the latter place for a few minutes, she aborted in the sixth month." Jansen, as quoted by Sand, reports the case of a cow from an aborting herd having been taken into a herd that had heon previously quite \'rcr from the disease. Soon after her arrival she aborted, and later cow after cow of the original herd aborted. The owner kept the matter a secret, and sent his cow to a neigh- bor's hull for service, with the result that for two years ahortion prevailed among cows served by this hull. McFadyean and Stock- man (3) later, in experimental work, attempted hut failed to in- cows by using a soiled bull for service. Hadley and Lothe (4) state; "A Large number of stockmen hold that the hull is an important factor in the transmission of contagious ahortion in herds. A smaller number believe that the hull merely acts as a passive carrier of the ahortion disease and is not actively concerned in the transmission." In a subsequenl bulletin, Hadley (5) re- marks: "The abortion organisms may enter the body during sexual intercourse." In an experiment carried on by the same author and co-workers, abortion-free virgin heifers were mated to abortion-infected bulls, infection being evidenced by positive, reactions to the complement fixation and agglutination tests. His results indicate, he believes, " that the hull is no1 so important a factor in transmitting ahortion as many believe." The con- clusions are: "Bulls may become infected with abortion- bacilli. Bulls thai reacted to the blood tests were incapable of dissemi- nating the abortion disease to the abortion-free heifers with which they were mated. Lulls appear to possess a sexual or individual immunity to ahortion infection thai renders them less susceptible than cows and induces a milder form of the disease. The resist- ance appears to be due to certain anatomic and physiologic differ- ences in their sexual organs which make them less favorable places for the growth of the abortion germs than those of the opposite sex." Buck, Creech, and Ladson (6) applied the agglutination test to 325 mature bulls, of which 288 were negative and 37 positive. Bacillus abortus was isolated from five animals, of which three showed marked lesions, two in the seminal vesicles, and one in the left testicle. They conclude: " B. abortus may involve organs of the generative apparatus of bulls, producing chronic inflammatory changes. Of the generative organs, the seminal vesicles appear to furnish the most favorable site for the lodgement and propagation of abortion infection." Schroeder and Cotton (7) cite the case of a bull which reacted to the abortion test and, on post mortem, Bad. abort urn was isolated from an abscess of one epididymis. They state : ' ' Our attempts to produce a similar case of infection artificially failed, and, in agreement with the difficulties many investigators have had to obtain incriminating evidence against bulls, we have thus far failed to infect bulls in any way that justifies the assumption that they are important factors in the dissemination of abortion disease." Further, they conclude: " Regarding the dissemination of abortion disease by bulls, we may say, however, that it would be foolhardy in the dim light of our present knowledge to take liberties with reacting bulls, or bulls from infected herds, or promiscuously used bulls." Cotton (8) failed to demonstrate the presence of abortion bacilli in the genital organs of the bull used to serve aborting cows, or in the testicles of two bull calves, one of which had been fed and the other injected with the cultures of the abortion bacillus. He con- cludes that the bull does not harbor the organisms in the testicles. Carpenter (9) injected both streptococci and Bad. abortum into the scrotal sacs of young calves, and intravenously in others. In no case was he able to recover the organisms from any part of the genital canal, except for a streptococcus in one instance. Rettger and White (10) were unable to obtain cvide-nce of the presence of Bad. abortum in three bulls slaughtered after repeated reactions to the complement fixation and agglutination tests. The three bulls had been under observation for three years, w r ith no conclu- sive evidence to indicate that they were a source of danger to the herds in which they were a part. They believe that the bull trans- mits the infection as a passive carrier. Attempts at artificial inoculation by natural channels have failed, with the possible exception of McFadyean, Sheather, and Minett (11) who were able to infect the bull by the prepuce in two cases and by the mouth in one case. The results, however, are by no means conclusive. They conclude, nevertheless, that cattle of any age of either sex may be infected by natural channels with the bacillus of epizootic abortion. 6 Schroeder (12) carried out investigations to ascertain the fre- quency with which hulls react to abortion tests, and the frequency with which lesions chargeable to abortion bacilli occur in the re- productive organs of reacting hulls, studies were also pursued which he states conclusively prove that hulls with infected repro- ductive organs may expel abortion bacilli with the seminal fluid. In the first two mentioned investigations 325 hulls from a Wash- ington abbatoir were tested, and slaughtered upon reaction. "Ap- proximately ten per cent of the hulls reacted, and approxi- mately ten pei- cent of the reacting hulls showed Lesions of the reproductive organs from which abortion bacilli were isolated.'* The value of these studies, he emphasizes, lies not in " that they give us a measure of the proportion of bulls that react positively to abortion tests or the proportion of reacting bulls that are car- riers of abortion bacilli," but in " the fact that they show that abortion bacillus disease of the hull's reproductive organs is not a wholly unique affection which practically may he ignored, hut an important condition that must he taken into account in our efforts to eomhat infectious abortion, since it has been proved to be asso- ciated with contamination of the seminal fluid." In discussing the method by which infected bulls transmit the organisms to eat t le, he believes that leakage of semen from the penis, or vaginas of cattle after service, contaminates the food which subsequently gains entrance to their digestive tracts. As the result of a series of experiments, he states: " the results fail to justify in the least degree the assumption that cows are infected with abortion bacilli via their vaginas or utcruses at the time of copulation, or that the hull, through copulation, is an agent in the spread of abortion disease." The work so far alluded to, has been limited to infection with, and the transmission of, Bait, abortum and the lesions associated with such infection. The last mentioned author, however, states: "A searcb for other specific causes of abortions among cattle has not. been neglected, and bureau investigators could relate at great length stories similar to those which investigators have told about microorganisms isolated from the products of abortions and the utcruses of cows that have aborted. Bacilli of various kinds, different types of micrococci, and spirilla or vibrio have been found repeatedly; hut when their pathogenicity has been tested in accord- ance wiih widely recognized and accepted and required bacterio- logical standards, not one shred of evidence has been obtained to prove them true etiological factors of bovine abortions. What role such microorganisms may have as causes of the sequellae of in- fectious abortions, and of other, possibly, independent, abnormal processes in the reproductive organs, is far from clear and merits careful study." Hadley (5) mentions the fact that: " Tuques- tionably the male often becomes infected with the germs that produce the various secondary diseases in the female, which are properly classed under the more inclusive term 'abortion dis- ' ing of the rarity with which the bull acquires abortion infection, he alludes to the fact that he may act as a " mechanical carrier of various disease germs from an infected to a healthy cow." Carpenter (9) working on the female genital tract, comes to the conclusion that, in all probability, the genital organs are normally free from bacteria. Barney (13) quoting Huet finds that bacteria may be present in the seminal vesicles of healthy animals (horses, cattle, pigs, and laboratory animals). This, he states, corresponds with the well recognized findings in other parts of the genito-urinary tract, not only in animals, but in man. He (Huet) has further found that in animals dying of acute septi- cemia, the specific organism (anthrax, pneumococcus) is to be found in the vesicular secretions. Furthermore it was definitely shown that an infection could be transmitted to the female during the act of copulation. Williams, W. L. (14), calls attention to the lack of vet- erinary literature relating to the pathology and bacteriology of the male genital tract, except as related to infection with Bad. abortum. Infection with other types of bacteria is empha- sized, the clinical recognition of such, with the accompanying pathological changes, and of the numerous phenomena involved in the process of reproduction in the male. The semen and its essen- tial germinal elements are taken up with reference to the entire lack of study devoted to them, and some of the abnormal changes are described. In a later contribution (15), he takes up the part played by the bull in the dissemination of genital infections and states: " Clinical studies now indicate with great clearness that the bull is an active spreader of that group of genital infections which cause sterility, abortion, and related phenomena." Williams, W. W. (16) studied the semen with reference to steril- ity, emphasizing the importance of its examination in the diag- nosis, giving methods for collecting samples, staining of sperms, and some of the abnormalities encountered. The work is funda- mental, and should be of great practical importance to all interested in the problem. In a later paper (IT), he brings out a more ex- tended discussion of the question. He concludes that the clinical examination is of vital importance, and that the efficiency of the semen depends not only upon its physical properties but upon the number of spermatozoa that are motile, the degree of motility, degree of oligospermia, and the percentage of imperfect spermato- zoa, either deformed or immature. Of forty bulls examined, he finds that twenty, or fifty per cent, showed lessened fertility, and others, aside from this, showed minor changes in the genital organs or semen. The same author subsequently takes up the sub- ject of reproduction from the viewpoint of both sexes, but empha- sizing infection in the male, and the frequency with which lowered vitality of the germinal cells occurs. Hopper (18) states: "A diseased bull may manifest non-fertility or decreased potency in different ways — by repeated service to apparently normal females without conception, by a high abortion rate in females that have been apparently normal, by characteristic infections following the 8 use of any particular sire, or by abnormalities in the breeding tracl noted by rectal <>r physical palpation." The observations of Williams (19) in a pure bred dairy herd bring out quite clearly the relation of the bull to the dissemination of genital infections. The hulls in this particular herd were ab- normal in many respects, as demonstrated by pathological changes in their genital organs, bacterial invasion of the parts, abnormali- ties of the semen and spermatozoa, and the probable transmission of infection to the females. Several of the sires from this herd furnished much of the material for the early basic work of this investigation. Since then the tracts of other sires have been worked upon with (piite similar or identical results. To summarize the work already done, most investigators have considered the hull as merely a mechanical carrier of i'xtd . (thor- ium infection, though all are more or less suspicious of his ability to become an active spreader. Schroeder, however, states that the organisms are eliminated with the semen, hut infection of the fe- male occurs secondarily through the digestive trad by contamina- tion of the food with the semen. Other investigators bring out fundamental points demonstrating the importance of other organ- isms than the Bans; baeillus and call attention to the need of a more thorough study of the anatomy, physiology, and pathology of the male genital tract. Any study of the genital organs must of necessity rest funda- mentally upon a thorough knowledge of the anatomy and phy- siology of those parts. Too few of us have stopped to consider these questions thoroughly, with the result that our ideas on the problem are more or less vague. It is much easier to understand why abnormal spermatozoa occur so frequently, or changes take place in the semen with death or weakening of the germinal ele- ments, if we realize or stop to consider the highly differential pro- cess of spermatogenesis, and the various structures which contrib- ute to the formation of the semen. We must come to realize that each part of the genital tract is essential to the normal funetionine- of the whole, and that the genital tract and reproduction are in turn dependent upon the proper functioning of the entire body. Walker (20) emphasizes the importance of a thorough kno\vle< of physiology in stating, "Although the subject of sterility has attracted the attention of the medical profession: and although much has been written on its causes and treatment, it cannot he claimed that the practical results obtained up to the present time are satisfactory, or that when consulted for sterility, the medical man of today can hold out to his patient much more hope of suc- cessful treatment than the medical man of fifty years ago. <>ur failure in this respect is in the main due to an ignorance of the physiology of reproduction." To bring out some of these points, the anatomy of the tract will be reviewed briefly, together with the physiology of reproduction, and the various factors which should be considered in a study of the problem. Anatomy and Physiology In origin and early development the ovary and testis are identi- cal. The gonad and mesonephros or primitive kidney are de- veloped from the urogenital fold. The gonad first forms as a medio-ventral thickening of the fold, which gradually expands until it becomes attached by a mere stalk. At first, the gland is made up merely of a superficial epithelial layer, and an inner epithelial mass, or epithelial nucleus. In the process of develop- ment, large primordial germ cells migrate from the entoderm of the future intestinal canal, and pass through the stalk to the gonad. In the case of the male gonad, seminiferous tubules are very difficult to make out in embryos smaller than 24 millimeters. Then they suddenly differentiate out as solid cords of germ cells, while the connective tissue grows in around them. These connective tissue sheaths unite at the center of the organ to form the anlage of the mediastinum testis. The testicular tubules unite and con- verge toward the hilus, there to meet the anlage of the rete. At the mesonephric end of the testis, the rete first appears as a col- lection of cells, differentiating out from the inner epithelial mass of the gonad. These cells gradually grow out to meet the collect- ing portions of the mesonephric tubules on the one hand, and the seminiferous tubules on the other. The rete is represented as cords of cells at first, which in forty millimeter embryos hollow out to form tubules. The mesonephros, or primitive kidney, early starts to degenerate cranio-caudally, — the tubules becoming separated into a cranial and caudal group. The collecting and secretory parts of the cranial group separate, the collecting tubules growing out to meet the rete with which they unite to form the efferent ductules of the epididymis. The caudal group of tubules is vestigial and becomes the paradidymis. The mesonephric duct becomes the vas deferens, connecting as it does with the tubules of the epididymis, and emptying into the urethra at Midler's tubercle or, as it later be- comes, the colli cuius seminalis. The seminal vesicles arise as hollow saccules from the dorsal wall of the mesonephric duct just as it empties into the urethra. The prostate develops as an outgrowth of the dorsal urethra just posterior to Midler's tubercle. The bulbo-urethral glands appear as solid, paired, epithelial outgrowths from the entoderm of the urogenital sinus. Midler's duct, at first a solid tube growing from the anterior part of the mesonephros, and ending at Midler's tubercle, becomes a hollow tube, and in the female forms the entire genital tract ex- cept for the gonad and the lower part of the vagina. In the male, the anterior part remains as the vestigial appendix testis, and the posterior part, as the vagina masculina. Ellenberger states, how- ever, that this embryonic structure is very seldom seen in the mature bull. The Male Ueproductivt Organs include the penis and testes, together with the excretory passages which conned the testes with the urethral canal. Those excretory ducts include the epididymis, v.is deferens, and seminal vesicles. Posterior to their termination in the urethra, there arc connected the ducts of the prostate gland and the bulbo-urethral or Cowper's glands. Testes: The testicles of the bull arc relatively large. Varying with the size and age of the animal they measure from fourteen to seventeen centimeters in length, including the epididymis, and from six to eight centimeters in diameter. Each testicle is en- closed within a serous sac. the tunica vaginalis, whose visceral layer is very intimately fused with the underly- ing covering of the organ, the tunica albuginea. The tunica albuginea is Quite thin and consists of connective tissue which is rich in elastic fibres. Muscular tissue is not present as it is in the case of many mammals. Inside the tunica, and closely attached to, though separated from, the parenchyma by a thin layer of connective tissue, is a layer of very loose connective tissue, which because of its rich supply of blood vessels is termed the tunica vasculosa. The parenchyma is of ;( yellowish gray color, and of a rather soft consistency. It is made up of the semi- niferous tubules, rete, and the connective tissue stroma, the medi- astinum testis. On section, the mediastinum appears as the center or axis of the entire organ. It is star-shaped, and radiates connect- ive tissue septa out into the parenchyma to support and separate the tubules. Ellenberger states that the testis of the bull and all ruminants lacks a closed system of interlobular septa, because of the feeble development of the connective tissue. The principal blood vessels and rete tubules are found in this Licture, the function of the latter being to connect the seminifer- ous tubules and the efferent tubules of the epididymis. The epithelium of the rete is quite irregular, -consisting in places of a single Layer; in others, of two Layers. At some points there are Formed groups of several cells lying over one another, with swollen homogeneous basal cells, which sometimes form projections into the Lumen. The interstitial tissue, besides conveying the blood vessels to the q, contains many " interstitial cells.'" These cells are rela- y sparse in the adult bull, and are comparatively delicate, slightly granular, often shuttle-shaped, with a rather small nucleus. Rmbryologically they arc derived From a syncitium arising from the mesothelium of the genital ridge, differentiating out by growth the cytoplasm. They contain Large quantities of fat. ami elaborate'lhe internal secretion of the testis. This secretion gov- erns the development of the secondary sexual characters, and has a profound influence on the general body metabolism, and develop- ■ I of the skeleton. The interstitial cells appear early in embryonic life even before there is any differentiation of sex, and 11 their greater relative development in the fetus is indicative of a future male development. In very young embryos, the growth is very rapid, followed, however, by a period of atrophy, during which the seminiferous tubules undergo marked development. Pende (21) states: " There seems to be an inverse relation be- tween the growth of the tubular and interstitial tissues, as one is hypoplastic when the other is in full activity." From birth to the onset of sexual maturity, which may be called a period of rest for the testicle, the cells are few in number. With the accentuation of the secondary sexual characters, and the beginning of sexual life, these cells again increase in number and activity. The parenchyma of the testis consists for the most part of the seminiferous tubules, which, on account of the courses they take in the different regions, are divided into groups. The peripheral tubules are the much-contorted tubuli contorti. These anastomose to form the much shorter tubuli recti. These in turn anastomose frequently, uniting to form the rete testis. The rete proceeds through the mediastinum to form the efferent ductules which break through the tunica albuginea to form the greater part of the head of the epididymis. The tubuli contorti are the longer and more numerous of the tubules, for it is here that practically all the spermatozoa are produced. The straight tubules are relatively so short that they may be regarded more in the light of the begin- ning of the system of excretory ducts. The seminiferous tubules consist of a thin peripheral membrana propria upon which rests the seminal epithelium, which is made up of the essential semen forming cells, and the cells of Sertoli. The spermatogenic cells may be divided into three groups, from within outward : the peripheral single layer of small cuboidal spermato- gonia ; one or two rows of large spermatocytes ; and three to five rows of spheroidal spermatids. The cells of Sertoli are more or less of the syneitial type, — large in size and irregular in outline. They occur at various intervals between the layers of spermatogenic cells, with their bases resting upon the membrana propria. Centrally they send out protoplasmic processes for variable distances, — some even reaching the border of the innermost cell layers. Spermatogenesis: In this process, the primary germinal cells, the spermatogonia, divide to form the primary spermatocytes. Maturation consists of two cell divisions of the primary sperma- tocytes, and these in turn form four spermatids. During the pro- cess, the number of chromosomes is reduced to half the number char? ct eristic of the species. The spermatids then become con- verted into mature spermatozoa. This mode of transformation may be seen in Plate I. In the process, the nucleus of the sperma- tid forms a large part of the head; the eentrosome divides, part passing to the extremities of the neck. One eentrosome becomes the anterior, and remains attached to the head, while the other divides to form the posterior eentrosome. The latter is divided into the anterior part, and the posterior nodule or annular ring. 12 Besides this, the posterior centrosome becomes elongated to form the axial filament, and the cytoplasm forms the sheaths of the neck and tail. The spiral filament of the connecting piece is derived from the cytoplasmic mitochondria. At this time, a large part of the cellular cytoplasm is east off. Meanwhile, the spermatozoa sink their heads into the Long protoplasmic processes of the Sertoli or " nurse " cells which furnish nutritive material for their com- plete development. Finally the adult cells are cast off into the lumen. The structure of the spermatozoa, and a discussion of the semen will he taken up Later. Epididymis: The epididymis is divided into three parts: the head. lio(l\ and tail. The head is made up principally of the lobules formed by the much -coiled efferent ductules proceeding from the rote. The ductules, ahout twelve in number in the bull, unite to form the body, which remains coiled and runs along the postero-medial part of the testicle to which it is more or less closely attached. To quote Ellenberger (23) : " The transition from the rete into the ductules is gradual, as the characteristic epithelium of the latter (ductules) begins in cavities without walls, and at first, gradually form a wall which is well marked out as a thin ring of interstitial tissue The epithelium of the ductules is in sharp contrast to the rete in t hat it has a single-layered ciliated columnar epithelium, in which here and there one finds round basal cells. The dark and light columnar cells alternate; the cilia are often cemented together, and form cone-shaped, homogeneous appearing protuberances. The secretory activity is quite clearly observable. In the light cells one finds secretory globules, accumu- lating in rows, sometimes above, other times below, arches of cells. The secretory droplets pass from the cells into the lumen, and often lie in irregular layers on the epithelium; also the 'nasal cells appear swollen and shoved out between the cylindrical cells." At the lower extremity of the testicle, the tail is formed, which is globular in shape, and more or less Loosely attached to the testicle. Here the ductules anastomose freely, gradually become less coiled, and end in the single excretory tube, the vas deferens. The epithe- lium at the tail part is more or less of the pseudo-stratified columnar, ciliated variety. Outside this is a memhrana propria, a circular muscular layer, and a connective tissue coat. The secre- tory activity is very marked here and one finds much secretion i'i the lumen. Courrier (21). working on the bat. suggests that the glandular activity is conditioned by the secretion from the inter- stitial (endocrine) gland. The action of the secretion is to dilute the large mass of spermatozoa present, nourish them to some extent and also stimulate them to active motility. Stigler (25) stales that the properties of the sperms are modified in the epididymis; the motility, the ability 1<> resist heat, and other properties are augmented, at least in the case of the guinea pig, rat. and mouse. Some authors state that the sperms firs) become motile when in contad with the prostatic secretion, but 1 have repeatedly exam- 13 ined the contents of the tail of the epididymis of the bull, rabbit, and guinea pig, finding full motility in each case, though the dura- tion is not nearly as long as when the sperms are ejaculated in the semen. The Vas Deferens is quite narrow (2 mm.) and runs from the tail of the epididymis to the verumontanum, or colliculus semin- alis, where it empties into the urethra in common with the duct of tlic vesicle. At first it is lined by epithelium similar to that of the vas epididymis, but this changes over into a peculiar low stratified type. Ellenberger describes it as follows: " The epithelium shows a very pronounced basal coat. The overlying cell zone shows more (at the most, three) rows lying over each other of elongated nuclei, while an outline of cell form is not ordinarily noticeable, so that it may be spoken of as a syncitium, and at the same time as a many layered epithelium." The mucosa forms low, broad folds into the lumen. The tunica propria is a thin con- nective tissue layer. The submucosa consists of thin connective tissue. Three muscular coats are present : an inner thin longitud- inal layer, middle circular layer, and an outer longitudinal layer. All are more or less intimately blended, and are surrounded by the adventitia, made up of connective tissue, elastic fibres and scattered longitudinal muscle cells of the internal cremaster mus- cle. Near the dorsal surface of the bladder, the ducts come in close apposition, and for ten to twelve centimeters dilate to form the ampullae. Here the mucous membrane bcomes much plicated, forming long folds which anastomose freely. The function of the vas is to convey the spermatozoa and secretions from the epididymis to the urethra. Disselhorst (26) believes the ampulla acts as a seminal reservoir and states that he has found spermatozoa stored up in the little pockets in the w r alls of this structure in animals during the rutting time. He suggests, further, that there is a relation between the state of development of the ampulla and the time occupied by copulation. When the organ is small or absent, as in dogs, cats, and boars, the coition is a slow process, but when the ampulla is large and well developed, as in horses and sheep, the coitus requires a relatively short time. Inasmuch as coitus is so rapid in the bull, and the ampulla is so well developed, it seems as though this function is very probable. The Seminal Vesicles are very compact glandular structures tying on either side of the median line, on the dorsal side of the •ladder, and ventral to the rectum. In the mature bull they tieasure ten to twelve centimeters in length, four centimeters in width, and about two and one-half to three centimeters in thick- uess. The glands are distinctly lobulated, quite tortuous, and are often asymmetrical in size and shape. They converge posteriorly, to empty into the urethra at the colliculus seminalis with the ampulla, in a slightly oval slit in the mucosa. Microscopically, the gland is of the anastomosing tubular type, with very pooriy 14 developed excretory ducts to the glandular cavities. Posteriorly one finds centrally a few sinus-like narrow excretory passages, which open into the somewhat Larger collecting and excretory duct. The epithelium is of the simple columnar type and produces a relatively Large amount of secretion. The gland cavities are sur- rounded by a membrana propria, over which is a relatively thick Layer of smooth muscle. < >utside this is a connective tissue covering which sends trabeculae or septa in between the Lobules. The secre- tion of the seminal vesicles is a tenacious albuminous fluid with a slightly yellowish tinge, all or pari of which appears in the ejaculate in the form of swollen sago-like grains which are soon dissolved following ejaculation and the liquefaction of the semen. The proteid compounds belong to the group of histones. The secretion is liquid when warm and coagulates when cold. Some say that the filling of the vesicles serves to excite sexual feeling, but this is doubtful in view of the fact that in seme animals the sexual desire exists before the vesicles are tilled. Likewise, Stein- ach found that rats, whose seminal vesicles had been removed, still retained their desire for copulation. The function of the secretion is to furnish much of the volume to the semen, and in some way ii has a. distinct bearing on fertility, inasmuch as ex- tirpation of the organs it: rats leads to lowered fertility. The vesicles of the bull are in mo sense a store-house Tor spermato as is usually understood. Repeated examinations in a Large num- ber of hulls have led to the finding of spermatozoa there only in very rare instances. That they serve as a resorption place for sperms thai are not ejaculated is also very unlikely. Normally, one - on smear of the vesicles, occasional cells, Leucocytes, lecithin gran- ules, sago bodies, and rarely a, few degenerated spermatozoa. The COLLICULUS SeMINALIS is a rounded or cone-shaped emi- nence in the posterior urethra, upon which the ducts of the sem- inal vesicles and vasa deferentia open. The duds open separately at the bottom of two narrow slits, one on each side of the mound. there being no distinct ejaculatory duet as in man. The function of the colliculus or verumonl anum is not definitely known. It is generally believed that the structure is made up of blood sp which become engorged during erection, causing a blockage lie- posterior urethra, which prevents regurgitation of the semen. Rytina (27), however, demonstrated that the structure is not ci posed of any unusual number of blood vessels or spaces, and that removal of the organ was ool followed by regurgitation of the semen into the bladder during ejaculation. He believe-, ami quite logically, thai its function is to afford a prominence upon which the ducts may empty. The mixture of the thick gelatinous semen with the thin prostatic secretion must occur at the moment of ejaculation and musi lie perfectly homogeneous, otherwise Large numbers of the organisms remain in the thick gelatinous portion of the fluid. The eminence serves this purpose in that the pros- tatic ducts which converge toward it. may ejed their secretion 15 toward the eminence, producing an admixture more evenly and quickly. Prostate: The Lull pos.ses.ses what Ellenberger calls a diffuse prostate. That is, there is no distinct glandular body as in man. It is composed principally of a glandular sheath around the urethral wall. Just posterior to the neck of the bladder, and in front of the urethral muscle, there is formed a slight dorsal trans- verse elevation, extending downward on the sides. This is what might be termed the body. The greater part of the gland is "disseminate" in form, being a sheath of glandular tissue em- bedded in the urethral wall. Dorsally it is about ten to twelve millimeters thick, and ventrally about two millimeters. The gland is a branched tubular structure, the interlobular tissue of which contains much unstriped muscle. The lobules are lined by a columnar type of epithelium. The ducts, about thirty to forty in number, open into the urethra in two rows posterior to the colliculus. The secretion is a thin, slightly turbid fluid, of a faintly alkaline reaction. Its function is to dilute the semen, stimulate the motility of the spermatozoa and nourish them. Fish (28) believes that the activating property of the secretion is due to enzymes, because boiling deprives the fluid of its power to accelerate the motility of the spermatozoa. Serrlach and Pares, quoted by Marshall (29), working on dogs, have adduced evidence indicating that the prostate is an internal secretory gland which controls the testicular functions, and regulates the process of ejaculation. It is stated that if the prostate is removed, sperma- tozoa are no longer produced in the testis, and that the secretory activity of the accessory genital glands ceases. These changes, however, can be prevented by the administration of extracts of the prostate. The fact that the prostate .elaborates a secretion having a definite relation to the testis, has been verified by other authors. Cowper's Glands (Bulbo-Urethral) : These glands are paired, oval structures about one and one-half by two and one-half centi- meters in size, situated on either side of the dorsal pelvic part of the urethra close to the ischial arch. They are deeply embedded, with the bulbus urethae. in the bulbo-cavernosus muscle, thereby being inaccessible to palpation. In general, they are of a well developed anastomosing tubular type. The connective tissue stroma is relatively thin, and thickens only in between the larger lobules, where one finds the larger collecting ducts. Each gland opens by a single duct. The epithelium is of the simple euLoidal type. Little is known of the function of its secretion, though Kingsbury (30) regards it in the light of a mating gland; that is. it lubricates the genital passages during coitus, as does its homologous structure in the female, Bartholin's gland. Ellenberger describes the urethra, slit ventrally, as presenting the following picture: "The colliculus seminalis distinctly ap- 16 pears as a process or offshoot of the crista urethralis of the Tri- gonium Vesicae. At the summit, and at the bottom of the two slits, open laterally the ducts of the vesicles, and medially the ductus deferens .... Prom the caudal slope of the coUiculus go two distinct mucous membrane folds which run through the entire pelvic urethra, aear together, somewhat diverging, and then coming together, so thai they form an elongated, narrow oval. At their caudal union, the excretory ducts of the bulbo-urethral glands Open side by side. Al the point of departure of the folds from the colliculus, arises a niche-shaped opening, between both folds, and likewise Lateral to each fold. In these niches open the ducts of the prostate. The openings of the disseminate prostate lie in rows as in the horse. bu1 form not less than six rows. There is b- ^•rvationes Anthonii L. de natis e semine genitali animaculis ( I'pon the formation of young from procreative material). The sperms were taken to be animals on account of their motility, and their significance remained questionable if not unknown. Spal- lanzi. quoted by Marshall (29), was the first to show that the filtered fluid was impotent, and that spermatozoa in the semen were essential to fertilization. KoUiker, in 1S41. discovered that the sperms arise from the cells of the testis, and Barry in 1843, observed the conjugation of sperm and ovum in the rabbit. This led to a clear understanding of the function of these important germinal elements. The spermatozoa are the male procreative cells, and are char- acterized by the possession of a head containing the chromoso necessary for fertilization, and a tail capable of propelling the organism on iis way to meet the ovum. The Length of the entire sperm, including the head, is seventy-five to eighty microns. The head is nine and live-tenths microns long, and five and five-tenths microns wide. It may be divided into two principal parts, the head and tail. The head, for the larger part, is made up of the nucleus, and may be differentiated by staining reactions into a darker staining posterior part, an anterior lighter part, and often a still lighter area between the two. On the anterior part is a sharpened edge, the acrosome, which serves to perforate the ovum. 'Idle whole is surrounded by a very definite limiting membrane which often becomes obscured under abnormal conditions. The tail may be divided into three parts: connecting piece, principal part, and terminal filament. The connecting piece, tin- essential motile apparatus, is the thickesl and strongest part, and joins the tail proper to the head. H consists of the central axial filament, a spiral filament around this, and an outer mitochondrial covering. Anteriorly it is limited by the anterior portion of the posterior chroi and posteriorly by the annular chromosome. The 19 anterior chromosome is directly connected with the head, there appearing a light break here at the neck where separation fre- quently occurs. This neck serves as a joint for the motion. The axial filament, therefore, does not reach the head, but extends back from the anterior part of the posterior chromosome. The principal part consists merely of the axial filament, and a thin outer covering, while the end piece is quite thin and is made up solely of the uncovered axial filament. The finer structures are seen only when special staining reactions are used, and then only when the sperms are obtained directly from the testicle. The function of the sperms is of course primarily that of fertilization. Numerous observers have, however, thought that they might have some other definite, though unknown, use. An editorial in the Journal of the American Medical Associa- tion (33) raises several important questions regarding this ob- scure phenomenon. The fact that an enormous number of sperma- tozoa are produced, and only one, or at most, a few perform the function of fertilization, raises the question as to what becomes of the remainder. It is stated: "Zoologists have found that in some of the invertebrates the spermatozoa invade the entire body of the female, and in some species they reach the ovum by pene- trating the cuticle from outside and migrating to their goal. Studies on rodents show that the sperms invade the epithelium of the generative mucosa and underlying connective tissue. These tissues seemed to be stimulated to growth, suggesting that this may influence the uterine mucosa in its preparation for receiving and embedding the egg, and in forming the decidua." It has been shown that the sperms contain a specific protein capable of producing antibodies in the blood plasma, by citing the fact that rabbits develop a distinct Aberhalden reaction for testicular pro- teins shortly after cohabitation. One very important observation showed that by immunizing female rabbits with sperms they were rendered sterile for some time, although after a few months they again became capable of impregnation. The question raised is : ". . . . if the spermatozoa invade the female tissues and cause the formation of specific antibodies which are capable of prevent- ing fertilization, may not such a process participate in the prob- lem of sterility?" This very problem seems to be a factor in ex- plaining why some couples who are not fertile to each other subsequently are both fertile when they cohabit with other indi- viduals. Motility: After clinical observation of the motility of the sper- matozoa of the bull, I find that it differs little or none from the types as observed by Reynolds (34) in his work on human sperma- tozoa. His observations are so accurate and well described that they will be given in his words. "All normal motions appear to be consecutive phases. Initial motion, i. e., motion as seen in fresh semen under favorable conditions, consists of a lashin? of the after part of the tail from side to side which is so rapid 120 as to constitute vibration. It produces rapid forward motion in a practically straighl Line, the head, middle piece, and forward por- tion pi' the tail maintaining their position in the line of motion with practically no swaying from side to side. The action of the flagellum is so rapid that it is quite impossible to follow its indi- vidual movements. Spermatozoa swi mm ing in this manner head against the current and usually cross the held of observation in about five seconds in the absence of currents or obstacles. This type of motion will be described throughout the paper as 'pro- gressive vibratile' motion. Progressive vibratile motion is nor- mally succeeded after a variable length of time by what I regard as the second phase of normal motion. "The second normal motion differs from the first not only in its character but in markedly reduced speed. The tail movement alters to a long stroke from side to side and almost the whole length of the fail partakes in the stroke. This is, moreover, accom- panied by swaying of the head and middle piece through an arc which is. always considerable and may even equal ninety degrees. The general outline of the spermatozoa, from being practically straight with almost non-detectable sharp, quick, small are vibra- tion of the aftertail, has become an S in outline, with large, slow, plainly perceptible undulations traveling gradually backward throughout the length of the spermatozoon. Speed has been lost and direction seems to be more specifically determined by the surroundings. Individuals a1 Ibis stage show a pronounced choice of direction and go up to objects in the medium, from which they later make off as though the movement were determined by tactile reaction to some extent. This type of motion has, therefore, been named 'undulatory tactile' in contradistinction to 'progressive vibratile.' 'The third type of normal motion succeeds the second and con- sists in a tendency on the part of the spermatozoon to push itself against or into any small masses of cells, or sometimes other materials, winch it may find in the neighborhood, bunting itself into any small cove that can lie found, and maintaining a slight burrowing motion by a lashing tail movement of the vibratile type no1 unlike the movements of the caudal fin of a fish. The move- ment of the flagellum in Ibis third type is unlike the second type in that it is vibratile rather than Lashing; but is slower than the vibratile motion of the firsl type and less limited to the after- part of 1he tail. These spermatozoa are apparently not caught in the debris or unable to move off. From time to time, they back out of such a cove and seek another mooring place. "This 'stationary bunting' motion is less universal than the other two. Many individual spermatozoa fail to attain it. It OS probable thai only the most vigorous individuals ever reach this st;i<_r<.. If has not been encountered in unmixed semen or in any artificial mediums, it has been observed only when the sper- matozoon is in the secretions of the female genital tract. It is 21 most frequent when the spermatozoon is in contact with a nest of epithelial cells .... "The three types of normal motion are not only distinctive but are always consecutive, i. e., the second follows the first after a period which is apparently determined both by initial vitality and by the favorable or unfavorable character of the medium, while the third has been observed to occur only in individuals which have already developed the second. They apparently constitute a normal cycle. "This cycle is open to the theoretical explanation that these types of motion are directly adapted to the function of the sper- matozoon ; thus, the progressive vibratile motion which is char-, acteristic of the earliest period of its existence appears especially suitable for its prolonged journey through the cervix and uterus to the fundus and tube. This is supported by the fact that during this motion it always heads directly against any existing current, and that during this stage of its journey it must under natural conditions be continuously exposed to the faint outward ciliary currents of the mucous membrane of these passages. "The undulatory tactile motion which succeeds the progressive vibratile would then be well adapted to the later stage in which the spermatozoon has reached the tube, where its success in conju- gation is dependent on its finding the ovum rather than on further progress. "The stationary bunting type of motion is that which would be demanded by the passage of the spermatozoon through the egg membrane which has been so often observed in the lower animals. This very plausible explanation is, however, necessarily theoretical and must always remain so, as the conditions which surround the specimen on the field of the microscope vary in so many respects from those in which it accomplishes conjugation in the course of nature; but the practical importance of the study of types of motion is not affected by their explanation." The duration of motility is a variable factor, dependent entirely upon the environment in which the spermatozoa are placed. Within the body they usually survive at least a week. One author describes a case in which he found living sperms in a woman who stated that coition had not been experienced for three and one- half weeks. It has been stated with regard to human sperms, that their motion should continue or be capable of being re-established for twelve hours. Cary (35) states, "first, that in their proper medium and at the body temperature the viability of the sperm cells may extend over a period of a few days; second, that their prolongpd vitality is probably dependent upon the normal lime salts of the prostatic fluid ; third, that the sustaining power of the seminal fluid is increased by its union with the normal secre- tion of the female genital tract." After death of the male animal they retain their motility in the genital tract for twenty-four to forty-eight hours.' 22 Wolf (36) worked on this problem in rabbits, and summarizes as follows: "The motility of rabbit spermatozoa can be preserved a1 Least nine days by placing the juice of the epididymis in the Tyrode solution which had been buffered and to which glucose had been added. The solution is adjusted to a pH of about 7.4. < >xygen is passed in and a suitable amount of sodium bicarbonate idded. The preparation must be kept at a temperature near the freezing point of water." Under ordinary conditions motility persists outside the body only a few hours after ejaculation, but if the semen is kept quite cool till the time of examination on a warm stage, motility should be capable of being restored in at least a fair percentage of the sperms for twenty-four to forty-eight hours. The cells are more sensitive to heat than to cold, and even to dilute acids more than to alkalies. Henle (quoted by Ellenberger) states that a spermatozoon under favorable conditions travels at the rate of twenty-seven millimeters in seven and one-half minutes, which makes three and five-tenths .millimeters per minute. This is about sixty times the entire length of the spermatozoon, and twenty-one centimeters in an hour. Forward motion is also more pronounced when the swim is against the current, such as is produced by the cilia of the oviduct. It has been demonstrated that feebly motile sperms become very actively motile when placed on the mucosa of a fresh Fallopian tube. Technique The material used in the work came from abattoir animals, hull calves and adult bulls raised in the department herd, and sires upon which clinical observations had been made by various veteri- narians in the field. Semen samples, many of which were sent in, were collected as often as possible after the method described by Williams (Ifi). The genital organs were removed with as little chance of contamination as possible, and taken or sent to the laboratory where the examinations were made soon after arrival. All cultures were made by searing the surface carefully, tearing nit a small portion of the tissue with sterile forceps, and placing it upon the media. In most cases, however, where fluids were present, tubes were inoculated with the material which had been drawn off with a sterile pipette. As stated by Carpenter (0), in Vis work on the female genital tract, the organisms usually live in the depths of the tissue. The media used principally were glu- cose glycerin agar (glucose 1 per cent, glycerin 3 per cent) ; plain agar, both with a pll value of 7.4, and hoeffler's blood serum. Small amounts of sterile blood serum or defibrinated blood were added to most of the agar slants to insure better growths of streptococci when present. All tubes, to which the serum had been added, were incubated for forty-eight hours before inocula- tion to insure absolute sterility. After inoculation, the agar tubes were sealed with sealing wax in give a partial oxygen tension which was quite necessary in iso- 23 lating the streptococci. The growth of other organisms was by no means hindered by the procedure, for one tube from eacii organ was often left unsealed. Incubation was at 37° C, and the routine method of examining the tubes was identical with the method of Carpenter (9). Whenever possible, a sample of blood was obtained from the animal either before, or at the time of slaughter, for agglutination with Bad. abortum antigen. Extracts from the seminal vesicles, testes, and epididymes were injected into the guinea pigs and examined at the end of four to six weeks for the presence of Bad. abortum. Sections of all organs were fixed as soon as possible in either Zenker's or Helly's fluid. Hematoxylin and eosin were used as routine tissue stains. Eosin and methylene blue, and Mallory's connective tissue stain were, however, frequently utilized for special staining reactions. The motility of the spermatozoa is best observed about half an hour after ejaculation, when the thick tenacious clot has started to liquefy. A drop of the fluid is placed upon a warmed slide, preferably one with a slight depression in it, and observation made with high or low powered objectives. The semen may be examined whole, or diluted with physiological saline solution. In the latter case, the sperms have a greater opportunity for freedom of motion in the absence of the thick viscid coagulate. A small vial of saline solution may be carried in one's pocket where it will be kept warm, and a drop of this placed upon the glass slide. If the clot of semen is merely touched to this drop on the slide, plenty of spermatozoa will be deposited for an examination. This method is very satisfactory for the observation of motility, but needless to say, the undiluted semen must be used for the deter- mination of the number of sperms present. If necessary, the specimen may be covered with a cover glass and the oil immersion objective used. While a warmed slide is quite sufficient to enable one to detect the presence of motility, the field soon cools and the sperms gradually become less motile. If possible, it is best to use a small electrically heated stage warmer, which keeps the field at a constant body temperature, so that the duration of the motion may be observed for hours if warmed physiological saline solution is added as the fluid evaporates. Stained preparations are best made with thin smears on the glass slides. This is conveniently clone by placing a drop of the semen on a slide and smearing it over the surface with the edge of another slide. A fairly thin and even field is thus obtained. A still better method is to first dilute the semen with physiological saline solution, so as to obtain fewer sperms in the field. After drying the preparation in air, fixation may be produced by draw- ing the slide through a gas flame several times, by immersion in equal parts of alcohol and ether, or even by the use of tissue Axel's such as Hellv's or Zenker's fluids. For ordinary staining:, heat '24 fixation is the quickest, and at the same time is quite satisfactory. After the slide is cooled, or washed, to remove the fixing solutions, it should be placed for a few minutes in a freshly prepared solu- tion (1 per rent) of chlorazene, as recommended by Williams, to remove the mucus and proteid material which otherwise blur the field, other authors (38) have recommended diluting the semen with about twenty volumes of a 0.12 per cent sodium carbonate solution in 0.8 per cent sodium chloride. From this liquid the eells should he centrifuged for several minutes, removed with a pipette, and smeared on the slide. Following this, the slide should lie thoroughly washed, preferably for ten minutes in running water, after which it is ready for staining. Numerous methods have been used for this procedure, but the sperms are more or less erratic in their reactions to the dyes, and one must be very careful to use the same method in all samples, in order to obtain uniform results. For quick staining, to bring out gross abnormal- ities of structure, and number of sperms present, one may use Gram's stain, or a light stain with any of the aniline dyes, such as fuchsin. To bring out the finer structure, particularly of the head, more careful teehnic must be employed. Carnetl and others (38) recommend the following: "The method o\' staining by iron-hematoxylin, particularly when supple- mented by a cytoplasm stain, has proved, on the whole, the most satisfactory, and possesses the additional advantage of being abso- lutely permanent, a quality that feAv anilines can boast. The method consisted of treating the fixed object — and here the fixing agent was heat — with a two per cent solution of iron-alum for from two to four hours. The excess of iron-alum was then com- pletely removed by pure water, and the object treated with a solu- tion of hematoxylin (one per cent aqueous) for twelve hours or Longer. The cells by this time were perfectly black. However, a 1 per cent solution of iron-alum removed the stain from the cyto- plasm, leaving the chromatin of the head, the centrosome, and the axial filament a brilliant blue-black. Care must lie taken that the preparation is not over-decolorized. After decolorization a satu- rated aqueous solution of eosin was added for from one to three minutes. This stained the protoplasmic envelope pink, and. unless the envelop<> is overstained. the view of the inner structures is not impaired in the least." Williams (17) recommends using two staining solutions, one of alcoholic eosin and fuchsin. the other a diluted methylene blue. The results obtained are. however, more or Less erratic, due to ih.' unstable character of the former stain, and the ease with which one may over or under slain. Many beautiful specimens may. nevertheless, be obtained by this method. 1 have frequently used ;i fairly quick method, though one not satisfactory in all s. which consists of staining for five or six minutes in a satu- rated aqueous solution of fuchsin. washing in water, and counter- staininsr for a few seconds in a strong solution of methylene blue. 25 A quite satisfactory method is to stain from two to five minutes in a saturated *aqueous solution of methyl green, with the application of gentle heat. The heat may be applied by warming the slide over a gas flame as it steams, or by placing the jar containing the stain in a hot water bath. The slide is then washed thoroughly and counterstained for five minutes in a strong aqueous solution of eosin. This is a fairly reliable method, and many excellent pre- parations may be obtained by its use. The nucleus is stained green, the anterior part of the head and all of the tail pink. So far, I have found this a very reliable stain for routine work. Pathology In the genital tracts that I have studied, a complete pathological and bacteriological examination was made wherever possible, but in many of the abattoir animals, and certain others, gross and microscopical examinations only could be made. The genital organs of one hundred and ninety-six males have been examined, and the gross or miscroscopical changes, or both, determined. The abattoir animals were from a large slaughter house, and a small local plant. Of the tracts, the pathology of which was studied, two were from aborted fetuses ; seven from apparently normal young calves ; four from mature fertile bulls; and sixteen from mature infertile or sterile sires. The remainder (167) of those examined were from abattoir animals. Besides these, three specimens of seminal vesicles, and seven of testes were studied. The tracts of the aborted fetuses and veal calves were apparently normal, both on gross and microscopic examination of the vesicles and testes. On gross examination, the tracts of the mature fertile bulls were normal, except for the presence of many fine connective tissue tufts and strands upon the serous covering of the tails of the epididymes, and adjacent portion of the parietal layer of the tunica vaginalis. Microscopic sections of all parts were apparently normal. The more important pathological changes in the tracts of the sixteen sterile or infertile bulls are given in the appended chart. The tracts are numbered the same as in Group VI of the report of the bacteriological findings; that is, any particular num- ber in either table refers to the same animal. References are made throughout the text to some cases which appear in this group of animals. Prostate and Cowper's glands are not included in the chart as they were not examined in some, and were negative in the others. Fibrous tufts and strands were present on the covering of the epididymes in each animal. The study of sections from the abattoir animals, as well as those from the sterile or infertile bulls, forms the basis for the follow- ing observations upon the pathology of the male genital tract. The tracts secured from the abattoir were studied for the most part on the basis of the organ rather than on that of the animal. For example, all sections of testes were placed in the same bottle of fixer, and the same plan followed for the other organs. 26 Testes: The testes seldom presented gross alterations of struc- ture except for abscess formation, which, according to Williams, occurs more frequently in the bull than in any of the other domesticated animals. Be also states thai arrest in development by which the organs remain soft, flaccid, and somewhat smaller than normal is not uncommon. One very interesting specimen, which typifies abscess formation, came from a bull witli a history reveal- ing that one testis had become much enlarged, hot, and painful. These symptoms developed very rapidly. Anorexia was well marked. Local applications were used for several weeks, but ;it the end of two months the condition was so little improved that unilateral castration was performed. The general condition of the animal soon improved, but after a year of service he was so uncer- tain compared to what he had been, that he was sent to the butcher. It was impossible to obtain the other testicle for study, though it undoubtedly was abnormal. The testicle removed was considerably enlarged, measuring twenty by ten and one-half centi- meters. The tunica albuginea presented a thickness of six milli- meters, and was made up <•;' firm selenitic tissue. The epididy- mis was not recognizable in the mass. Testicular tissue was almost entirely gone. The only remains, of what appeared to have been parenchyma, was an elongated irregular area at one side of the organ. This tissue consisted of a whitish opalescent material, speckled with varying sized abscesses. This organ is pictured in Fig. 3. The remainder of the organ consisted of a thick yellowish caseous mass. Streptococcus viridans was recovered from the outer portion of the organ, and guinea pig inoculations failed to demonstrate Jiact. abortum. Microscopically, changes are quite common and varied in char- acter. In the seminiferous tubules, the changes range from a slight desquamation of the germinal epithelium to atrophy and complete degeneration of the entire tubule, as was the ease in the Lefl testis of Bull 1. In the mild cases, spermatogenesis occurs apparently in a normal manner up to the spermatid stage, at which point many of the cells degenerate and slough off. These appear in the seminal fluid, associated with the few sperms that reach maturity. This sloughing and degeneration may be localized in a few of the tubules, or it may be widespread over the entire organ. I ikewise, the changes may involve not only the more mature cells, but they may be so severe as to cause almost total degeneration and desquamation of the seminal epithelium, as in Pig. !•">. These defects in spermatogenesis are of course evidenced in the semen by the presence of immature, or abnormal types of sperms. With cessation of spermatogenesis or degeneration of the epithelium of the entire eland, no sperms are formed. Not infre- quently one finds numerous tubules, or even the entire testis in which the germinal epithelium is intact, but there is little or no evidence of mitosis, as in some tubules of Bull 0. The cells are several layers deep as in the normal condition, but they are not dividing. This condition is shown in Fig. 13. 27 In the more chronic forms, the tubules become atrophied, and frequently disappear entirely. The membrana propria may lie- come thickened, due to excessive connective tissue formation, or infiltration with, serum or exudate. On the other hand, a dis- tinct atrophy may occur. The stroma of the organ not infre- quently is thickened by inflammatory exudates, or by a noticeable increase in the connective tissue. In some testes, the connective tissue is so much increased that the tubules rapidly become atrophied, and disappear. In abscess formation, due to acute in- flammations, the entire organ becomes enlarged, markedly hyper- aemic, and infiltrated with leucocytes. Necrotic areas appear here and there in the parenchyma. The rete often shows a marked de- generation of the lining epithelium, and atrophy caused by in- crease of the interstitial connective tissue. Epididymis: This organ not infrequently presents gross ab- normalities, and very often is pathological on microscopic examina- tion. Acute inflammation, with induration or abscess formation, is very common in the tail, but less so in the head and body. Possibly this is caused by the fact that the tail is the most pendant portion of the organ. In these cases, the tail is enlarged, soft, and quite hot and painful on physical examination. Enlargement, due to a connective tissue induration, occurs occasionally in all three parts, and the inflammation may produce adhesions to the adjacent serous membranes. Inflammation of both the parietal and visceral layers of the tunica vaginalis is very common. In those cases, the membrane usually is quite hyperaemic, and on the surface it presents many small reddened tufts of newly-formed connective tis- sue. In adult bulls it is exceptional not to find at least slight evidence of some previous inflammation. In all of the numerous bulls ex- amined, both apparently normal and sterile, I have found but one in which some evidence of inflammation (either present or past) could not be found. Along with the fibrous tufts, are numerous fine strands of connective tissues passing from the covering of the tail of the epididymis to the adjacent portion of the parietal layer of the tunica. The strands often extend even to the upper part of the head. Microscopically, inflammation of the part is shown by hyper- aemia, loss of cilia of the lining cells, and exudation. In the more severe forms, the lining cells which furnish considerable secretion for the nourishment and stimulation of the sperms, become de- generated, and are exfoliated into the lumen, as in Fig. 21. This condition is very common in sterile bulls, and those of lowered fertility. In the chronic types, the interstitial connective tissue is increased in amount, leading to degeneration and atrophy of part or all of the tubules, as in the case of Bull 2. Infiltration with leucocytes, and necrosis, are the predominating lesions in the pyo- genic types of inflammation. 28 Dions Dkfekkxs: This tube seems to be peculiarly free from severe inflammatory processes, and when these appear they are limited to the mucosa. The cells of the lining membrane not m- 'ivijiitMitly show a mild type of degeneration and exfoliation, or in i be more chronic forms, the entire membrane degenerates and disap- pears. In man. the duct occasionally becomes occluded, but so far I have failed to find this condition in the bull. Undoubtedly, when the occlusion docs occur, it is near the origin of the duct at the tail of the epididymis. Seminal Vesicles: The seminal vesicles and epididymis, especially the tail, seem to be the parts most subject to extensive pathological changes, and bacterial invasion. In most instances, diseased vesicles present gross manifestations recognizable on clinical examination, while on the other hand microscopic changes may be present in the absence of gross lesions. As diagnosed on physical examination, or even on post mortem examination of th e tract, the various forms may be classified into: 1. Acute Catarrhal Type: In this form, the vesicles are usually enlarged, soft, and more or less reddened by hyperaemia. On physical examination of the animal, distinct flinching is produced when pressure is applied to the organ. Enlargement may even be absent in the early stages, and the diagnosis may be made from the extreme sensitiveness alone. 2. Suppurative or Cystic Types: In both of these types, the vesicles are usually enlarged, either uniformly, or, as is usually the case, in localized areas. The suppurative form may extend over the entire gland, forming one large encapsulated abscess, or on the other hand, it may take the form of variable sized abscesses with thick sclerotic, or thin fluctuating walls. < >eeasicnafly the abscess^ rupture and discharge their contents into the rectum. Dr. Wil- liams presented one case of this type. One vesicle was apparently normal, whereas the other was about five times larger than normal, and consisted of a dense outer capsule which was adherent to all surrounding parts. Oil dissection, it was found that the organ consisted of abscesses of various sizes, the Larger one of which had ruptured some time previously into the rectum, leaving the dis- tinct remains of an opening into that part. The cystic form may occur either with or without suppuration. One case came to my attention in which both vesicles were made up of abscesses of varying sizes as well as of a smaller number of cysts. Evidently the cysts were of the retention type, and were secondary to the pyogenic infection. 3. Chronic or Sclerotic Type: This form is characterized by a distinct firmness with or without marked enlargement. The con- dition may be accompanied by disease of the parenchymatous tissue or it may take the form of a chronic productive Inflammation of the interstitial tissue. This inflammation may be simply a super- ficial thickening, or it may extend in between the lobules. 29 4. The Peri-vesicular or " pan-inflammatory" Type usually is the result of severe inflammation of the vesicles, with probable rup- ture of some of the smaller cysts or abscesses upon the surface. The vesicles are, as a rule, considerably enlarged and buried in a dense mass of adhesions which involve neighboring structures. The vesicles cannot be palpated on physical examination, and it is only on careful post mortem dissection that they may be studied. This type, however, is quite rare, — two cases only having come to my attention. In both, the vesicles themselves were markedly affected. Microscopically, changes in the vesicles are quite frequently en- countered, even in the absence of gross manifestations. In the acute catarrhal forms, the mucosa and submucosa are hyperaemic. The lining cells show various forms of degeneration, and there are, as a rule, inflammatory exudates in the lumen. As the in- flammation progresses, the lining cells degenerate further, and become cast off into the lumen of the glandular cavities, as in Plate VI. The normal clear mucous secretion becomes mixed with fibrin, leucocytes, and cellular debris. These changes may involve merely parts of the organ, or they may be quite extensive With large sections, one may find the inflammation in all stages, from the mildest catarrhal type, to complete degeneration and exfoliation of the secretion-forming mucosal cells, and filling of the cavities with degenerated cells, leucocytes, and debris. Frequently the inter- stitial tissue is in no way affected, but at times it is thickened by oedematous exudates, leucocytes, and fibrin. The chronic inter- stitial form is characterized by a considerable increase of con- nective tissue, — producing marked atrophy, or even complete obliteration of the glandular cavities. Microscopically the sup- purative form may be diffuse over the entire gland, or as stated previously, may be in the form of localized abscesses, with or with- out a thick connective tissue wall. The parenchyma in these cases is usually extensively degenerated and atrophied in those parts that have not undergone suppuration and necrosis. The cysts appear to be of the ordinary retention type, and may or may not be accompanied by extensive changes in the lining epithelium. Both the abscess formation and cystic conditions are undoubt- edly initiated by an obstructive inflammation of all or part of the excretory duct. This is, however, a protective mechanism, for where the duct is closed the bacteria and exudates are unable to reach the urethra and contaminate the semen. Prostate and Cowper's Glands: These glands were more or less neglected in the early part of the work, but later were sub- jected to the same examination as other parts. Of the thirty-six of each type of gland examined, I failed to find one with any gross changes, but two prostates were found that presented a mild catarrhal inflammation of the mucosa. It is probable that Cow- per's glands, as well, occasionally undergo inflammatory changes. 30 Semen*: The semen, made up as it is of mixed products of the testes and accessory sexual glands, is very often abnormal, as would be expected in view of the frequency with which changes occur in the glands contributing to its formation. The normal semen is remarkably adapted to its function of nourishment and stimulation of the spermatozoa, and their conveyance to the in- ternal female genital organs. The spermatozoa are "extremely sensitive to changes in their environment, with the result that any alteration of the physical or biochemical content of the seminal fluid may cause death of the sperms. AVith this in view, we must remember that disease of any of the contributing organs is a poten- tial danger, and threatens the potency of the animal. Each or all of the glands may add bacteria, acid secretions, or inflammatory exudates. On the other hand, they may not function at all. In each case, however, the semen is altered. Unfortunately it is impossible with present methods to obtain the fluid absolutely free from vaginal mucus, but with care it may be secured reasonably free from contamination by douching the prepuce of the bull and vagina of the cow before service. Tins method was used as often as possible in collecting the samples. The usual amount of semen obtained was from six to ten cubic centimeters. With a hypersecretion of one or all of the glands, the semen becomes quite thin and watery, with a deficiency of solid matter, together with changes in reaction. On the other hand, hypofunc- tion results in a secretion too viscid, which is equally unsuited to the requirements of the spermatozoa. The thin watery semen clots imperfectly or not at all, and clotting is essential in protect- ing the spermatozoa from the acid secretions of the vagina. Likewise, a medium too viscid is a distinct hindrance to motility. Changes in reaction are very frequently encountered. The sperms are very sensitive to dilute acids, so that with even a slight acidity motility may diminish or entirely cease. Purulent inflammatory exudates are occasionally mixed with the semen, and although the pus cells themselves have not been found to be destructive to the sperms, certain degeneration products in the exudate are very toxic, and inhibit or destroy the motility. So far, I have failed to find red corpuscles present. One very interesting sample of seme?] was quite thick, of a yellowish green color, and of a dis- tinctly acid read ion. The secret ion from the vesicles was later found to be of this same character, and was due to a Pa. pi!o<>>fn- iwus infection. The vesicles were highly Inflamed and degenerated. The spermatozoa were in this ease markedly decreased in number. ami devoid of motility. The early precipitation <>:' the ''Boettcherchen" crystals seems to he intimately connected with sterile semen, or spermatozoi lowered vitality. Likewise, a decrease in solid matter is often - in a defii n. Fn normal semen, the clot disappears after standing a time, and a thick sediment settles out. This sediment is decreased in amount as a rule in abnormal semen. 31 Spermatozoa : Spermatozoa, the essential germinal elements, are very frequently abnormal, changes in which may be manifested in many ways. We may divide the deviations into changes in structure, and changes in the motility which is so indicative of the intrinsic vitality of the sperm. Reynolds (34) describes two forms of abnormal motion. The first is " rotary swim- ming," in which the sperms move forward progressively, and sometimes with fair rapidity, but in a spiral screwlike manner. He states that this type of swimming is very awkward, easy to recog- nize, and is usually of quite long duration. The other form termed "pendulum swimming," he states, is less common than the rotary swimming and is usually confined to relatively fewer sperms in a given field. "In this the middle piece and upper tail seem to lose their flexibility and balance to a considerable degree, and the loAver tail motion swings the forward part of the spermatozoon to and fro with a pendulum movement. This type of swimming yields very poor progress." One factor we must bear in mind in the study of the semen ob- tained from the vagina, is that the spermatozoa may be highly motile before ejaculation, but the admixture of hostile vaginal mucus may inhibit or destroy the motility. On the other hand, the conditions may be reversed. Cary (35), in one instance, found that the spermatozoa in a sample of semen collected from a condom, appeared to be of very low vitality, while when they were mixed with the vaginal secretions, an exaggerated activity was manifested. May we not have to contend with this factor in some herds in which there is a very distinct acid and toxic vaginal secretion from the products of cervicitis and vaginitis? In a study of motility, we must consider not only the abnormal types which may be encountered but the percentage of motile cells, and the duration of the movement. In necrospermia all the ejaculated cells are motionless or dead. In other specimens, vary- ing percentages of the cells are without motion, and the others may be possessed of full and lasting motility. On the other hand, the motility in some cases is very active at first, but quickly subsides even under the best of conditions. The appearance in freshly ejaculated semen of numerous sperms that have a tendency to take on the "undulatory tactile" type of motion when they should be in a highly active state, is very indicative of lowered vitality. Many specimens present this very picture, whereas the very active pro- gressive movement should, under proper conditions, survive for a considerable time before it gives way to the second, and slower type. The cells frequently early hunt into epithelial cells or clumps of immotile sperms, then back out and move around sluggishly, only to repeat the same performance till they stop moving en- tirely. I have seen one specimen in which the sperms all tended to clump. Whether this was the result of some agglutinative sub- stance in the vaginal secretion is problematical. I have seen several specimens of semen in which practically all the sperms were motile 32 when lii-st examined, but the motion did not survive for any great length of time. Even a small percentage of motionless sperms or of those showing lowered vitality is a considerable factor in potency. Although millions of the germinal elements are ejacu- lated into the vagina, large numbers of them are destroyed or be- come motionless there, and a small number is left behind in the cervix and uterus; so that even though but a single sperm is required for fertilization, the chances of impregnation are dimin- ished in proportion to the number of dead or defective sperms. Aspermia: Absence of spermatozoa in the semen is rarely en- countered, and is probably due either to total cessation of sperma- togenesis, or to an obstruction at some point in the system of excretory ducts. I have seen but one case of this character. The semen of this bull was greatly increased in amount, and of a thin watery consistency. Due to lack of cooperation on the part of the owners, the tract could not be obtained for study. Oligospermia, or a diminution of the number of spermatozoa, is quite common, and is undoubtedly associated with defective spermatogenesis, either as a result of poor mitosis of the seminal epithelium, or degeneration of the elements before maturity. This condition may vary from the finding of only occasional dead sperms in the field, to but a slight decrease in the usual number of normal sperms observed. Abnormalities in morphology may be classified into immature types, and deformities or imperfect development of the head and tail. Defective spermatogenesis occurs so frequently that it is not surprising to find spermatozoa in various stages of develop- ment cast into the excretory ducts. The various stages passed through in the development, from spermatogonia to adult sperm, are numerous, and it therefore is to be expected that we should see in abnormal semen many different immature forms. No classification of the various types can be made, hut a clearer understanding of them can best be obtained by a review 7 of the process of spermato- genesis. Spermatocytes and spermatids are seen in the more severe types of defective spermatogenesis, and are relatively uncommon, while the more mature forms that result from the transformations of spermatid to adult cell are very often seen. Rome of these inter- mediate types are large oval cells without distinct nuclei and as a rule with poorly developed tails. Cells with no tails or distinct nuclei, thos<> with protoplasmic appendages to the head or tail, and various other types, are occasionally encountered. Most of these are motionless and incapable of pfoducing impregnation. Others are active, but survive a comparatively short time. According to Cary, the production of the immature cells is- an effort on the part of the testes to sunplv an abnormal demand, and their presence indicates that the fertility of the semen is impaired. The deformities, which may be divided into cephalic and caudal groups, are also the product of defective spermatogenesis, or they 33 represent a degenerative process induced possibly by abnormali- ties of the fluid environment. It is rather difficult, however, to distinguish between deformities and immature types. The two most common cephalic deformities are what might be called macro and microcephalic forms. In the former, the head is enlarged to a greater or less extent, it is usually defective in staining qualities, and its outline is indistinct, due to degeneration of the covering membrane. This type is seen in Fig. 32. Also the shape of the head is usually abnormal, being either quite rounded, long and narrow, or short and very broad. Cells with protoplasmic ap- pendages, though they are more properly an immature type, occasionally give the head a greater volume. Microcephalic sperms vary from those slightly smaller than normal to those in which the head is represented by a slight knob. In some cells, the head is small and round, in others, short and stubby, while another type is normal in outline but diminutive in size. These forms likewise are, as a rule, deficient in staining qualities, and are undoubtedly degeneration forms, occurring either as the result of faulty de- velopment, or degeneration subsequent to their formation. Cary believes they are degeneration types because in the majority of cells the tail is apparently fully formed, and in the normal process of evolution the tail is the last part of the cell to be exhibited. Double headed forms are quite rare, but they nevertheless appear at times. Their significance is difficult to explain. Another very frequent deformity of the head is a marked constriction at the posterior part so that it is the shape of a pear or top as in Fig. 27. In some, the head is otherwise normal in size, while in others it is much elongated, as in Fig. 28, or considerably atrophied. A con- striction at the middle of the head, as in Fig. 29, is not uncommon. Both defects are undoubtedly the result of nuclear deficiency, as the nuclear part of the head in these cases is much diminished in size, and stains very deeply or not at all. I have seen spermatozoa, the heads of which were like an inverted cone, with a bulging rounded base. Other heads are even somewhat contorted and bent on themselves, as shown to some extent in Fig. 26. Under caudal deformities, the most frequent form encountered is a thickening of the connecting piece. This may occur as a uni- form thickening, or as a bulging appendage. Kudimentary de- velopment of the tail, the presence of two poorly formed tails, and defective development of the connecting piece occur rather infre- quently. All these immature and defective types are, as a rule, motion- less, and of course incapable of producing fertilization. Their presence indicates lowered fertility of the semen. Besides these deformities, there are sperms showing a curvature of the tail at an acute angle just posterior to the neck, — the so-called " wry neck." Their significance is difficult to explain, but they occur frequently in semen fixed and stained by the same routine methods used on samples in which thev are absent. They probably are not the re- 3 suit of the methods used in fixing and staining. Some think they are slightly immature types, or thai the condition is produced by abnormal contractions of the tail. The majority m sperms, how- ever, especially those from highly fertile bulls, do not show this y The mosl common changes in the spermatozoa, arc those in which there is a separation of the head from the tail, and degem lu ,n «»f the head as evidenced by reaction to stains. The separation of the head from the tail always occurs at the neck, and often is associated with degeneration or abnormalities of the head, rhe separation, in the majority of cases, indicates sonic Lowering vitality in the elements, although in many instances traumatism produced in making smears or collecting the samples is responsible. Various forms of abnormal staining of the head arc very common. The cell membrane, which is normally distinct ami sharp, becomes blurred in outline. Normally, the head takes a good differential .in in the anterior part staining Lightly, and the posterior part somewhat deeper. The nucleus is distinct in outline and well de- fined. Tin' lighter " inner body " stands out in well stained speci- mens. As the result of degeneration, the whole licad may take the stain uniformly, either slightly or much deeper than normal. ording to the degree of degeneration. The whole problem oi staining, however, depends very much upon the methods used. and the rare with which they are applied. When a good method is obtained, it should be adhered to. and used uniformly on all specimens. As a rule, however, a certain amount of practice will enable one to differentiate between the sharply outlined, clearly staining normal forms, and those that show abnormal reactions to the stains. Bacteriology A complete bacteriological study was made of the genital tracts of fourteen normal young veal calves (six to twelve weeks old), tour mature fertile bulls, and sixteen mature bulls, either sterile or impotenl to some degree. Together with these, the tracts of eleven aborted fetuses, seven calves dying from calf infections scours or pneumonia), and sixteen bulls slaughtered at an abattoir were studied bacteriologically. Occasionally, studies were made of indi- vidual seminal vesicle's or testes, when these parts alone were broughl or sent in. The history of the abattoir animals was. , -nurse, quite indefinite or entirely negative. On the killing floor. many tracts could be studied for pathological changes, but in the bacteriological work it was difficult to care for more than two tracts on each visit. The results of the bacteriological examinations are given in the appended tables, in which the tracts are divided into six groups. The results in Group T. consisting of normal veal calves, indicate that the genital organs of young male calves are. under normal conditions, free from bacteria. Carpenter (9) obtained like re- sults in examining the genital tracts of heifer calves. The cul- 35 tures made from the seminal vesicles and testes of all these veal calves were, with two exceptions, negative. Both seminal vesicles of one tract and one of another yielded cultures of Staphylococcus albus. Adult bulls of known fertility were naturally difficult to obtain only the four animals in Group II being- available for examination* Iwo of these (Nos. 1 and 2) were from the experimental herd kept by the department, and at all times had a good breeding history. The other two were good breeders, but were slaughtered because of poor pedigrees. Bull 1, raised in the department herd had a severe attack of scours when a few weeks old, while the calf- hood history of the other is not known, he having been purchased after reaching sexual maturity. The cultures from the «enital organs of the former (Bull 1) were entirely negative, except those from the left epididymis and scrotal sac. which vielded growths of Streptococcus viridans. All the organs oi ! the tract from this ani- mal were normal, except for the fact that numerous strands of connective tissue extended from the serous covering of the tail of both epididymes to the adjacent part of the parietal layer of the tunica. The tract of the other failed to show any organisms. The only evidence of any abnormality was the presence" 5 of the same connective tissue strands on the tail of the epididymis, as in the first tract. The other two bulls gave negative cultures from all parts. Of the sixteen bulls in Group III. slaughtered at abattoirs and m which no history was available, eight failed to show the pres- ence of any organisms in their genitalia. Of the others, the vesicles yielded cultures of Staphylococcus albus nine times, and strepto- cocci four times. Staphylococcus albus was recovered once from the prostate, and once from Cowper's glands. The testes gave cul- ■ tures of staphylococci in two cases, and Bad. abort urn in one. No observable anatomical changes accompanied the presence of the Bang bacillus in this case. The epididymes showed growths of staphylococci five times, and streptococci on three ° occasions Streptococci were isolated from the scrotal sacs of eight testes' The results in Group IV (aborted fetuses) show that bacteria are often present in the seminal vesicles or testes of these animals. As a rule, however, the organisms are identical with those isolated from the blood or other parts of the animal. This is to be ex- pected, however, for because of the feeble resistance of the fetus to any infection, the organisms circulating in the blood may be isolated, as a rule, from many different organs and tissues. * All samples of blood set with Bart, abortion antigen were negative, irrespective of whether or not the organism was recoverecf f rom the blood or other tissues. This is in accordance with the findings of Carpenter in the female fetus.— the resistance is so feeble that few or no antibodies are formed to combat any existing infection. Bact. abortwn was recovered in two cases from the vesicles, and in four cases from the testes, but in each instance the same organism was present in the blood or other tissues of the body. 36 The results from the tracts of the calves dying of calf infec- tions are given ... Group V. and show thai five were negative, lhe other two showed />'. coli, staphylococci, and streptococci, in the ; M s indicated by the chart. 1„ (i,, )U p VI. the mature infertile or sterile bulls, there was a comparatively wide variation in the type of organisms encoun- tered hut streptococci and micrococci were the most common in- vaders. In the order of the frequency of infection, the organs w . ollM be enumerated as follows: Vesicles epididymis (usually the tail), scrota] sac, testes, prostate, and Cowper stands, lhe firsl three parts mentioned usually contained bacteria. A strep- tococcus was the usual invader of the scrotal sac, and very prob- ablv was the cause of the connective tissue lulls and strands so frequently seen. The vesicles and epididymes gave, in the order of the frequency of their occurrence, staphylococci, streptococci B coli and Ps pyocyaneus. The streptococci were usually ot the viridans group, though a few were hemolytic and two strains were indifferent to blood. The testes gave growths in only eleven instances,— staphylococci eighl times, .streptococci two times and !ui unidentified rod once The prostate yielded staphylococci twice and ('owner's gland once. „ , .,._ \s emphasized previously, the vesicles and tail of the epididymis are most subject to infection and degenerative changes. At the same time, they are intimately connected with the secretion of the semen Once the epididymis becomes infected, there is nothing t () prevent the organisms, together with inflammatory products, from beim- mixed with the semen and ejaculated during coitus. Uso i" the vesicles, unless the inflammation is so severe as to occlude .he excretory duct, the organisms are mixed with tne .vesicular secretion, which is emptied into the urethra during ejaculation. Carried along with the bacteria, are, of course toxic products, degenerated cells, and the otherwise altered secretion of the elands. One interesting case noted was that of a bull that had passed from a state of fertility to that of complete sterility (lul . u „. a period of two months. The semen was semi-fluid, green- ill, ydlow in color, and contained a very few non-motile sperma- o oa Post mortem examination snowed that the vesicles had undergone abscess formation and that they contained yellowish Seen material similar to that which had been discharged during copulation Streptococcus hemohjtieus and Ps. pyocyaneus were isolated from both vesicles, and from the semen. Micrococcus albus was isolated in nearly all cases of vesiculitis and was otten asso- ciated with Streptococcus viridans or hemolyttcus. Bacteriological stuT.es of the semen are. on the whole, more or loss unsatisfactory, due to the present difficulty in obtaining sam- g free from any chance of contamination. In most ot the ab- normal bulls bacteria of various types were .sloated from the " m most of which agreed culturally with those later isolated from the internal genital organs of the same tracts The method , lf culturing consisted of douching the prepuce ot the bull and 37 vagina of the female with sterile saline solution before breeding. Samples of vaginal mucus were taken before service, and the flora compared to that after douching. This method of douching pro- duced vaginal samples relatively free from bacteria, at least so much so that the post coital fluid demonstrated that many organ- isms must have been introduced from without. "Whether or not they came in with the semen is problematical, but in all probability this was the method of introduction. I have so far failed to obtain Bad. abort urn from the tract of an adult animal, either by direct culture or guinea pig inoculation, except from the testicle of one abattoir bull. The agglutination tests with Bad. abortum antigen were all negative, except for two abattoir bulls. The results so far obtained would seem to indicate that, in accordance with the findings of other workers, the Bang organism seldom invades the male genital tract, or does not thrive there after its introduction. Schroeder (12) and others, have, however, on various occasions, recovered the organism from the bull, and the former author even states that it invades the vesicles and is eliminated with the semen. Discussion A complete discussion of those factors which have a bearing on reproduction and fertility in an animal, includes not only a thorough study of the genital tract, but an appreciative considera- tion of various extrinsic factors. The effect of environment has long been known to have a marked influence upon breeding, par- ticularly with reference to animals in domestication. Diet, though long relegated to a minor phase of the question, has, within recent years, come to be a matter of prime importance with regard to its bearing upon the entire body metabolism. The endocrine organs preside over and regulate the growth and functioning of the geni- tal organs from the earliest embryonic stage to the cessation of sexual life. Any derangement in one results in functional or organic changes in the other. In a given mating, Ave must take into consideration such factors as impediments to coitus, as well as those numerous agencies in the female which may interfere with the union of sperm and ovum, or with the successful implantation of the fertilized egg in the uterus, and its growth and develop- ment there till normal parturition takes place. Successful repro- duction depends upon the mating of sexually sound females to equally sound males. Considering the various factors which gov- ern reproduction, sexual soundness must necessarily depend, to a large extent, upon a good general condition of the entire body. Environment: The effect of environment on fertility in the bull is no doubt a minor factor. Cases in which changes in en- vironment affect fertility probably occur, however, particularly when fear and other psychic disturbances play a part. Marshill (29) states: "It would seem probable that failure to breed among animals in a strange environment is due not, as has been suggested, to any toxic influence on the organs of generation, but to the same causes as those which restrict breeding in a state of nature to certain particular seasons, and thai the sexual instinct can only be called into play in response to certain stimuli, — the existence of which depends to a large extent upon appropriate seasonal and climatic changes." Diet: Under this heading we may include not only the effect of deficient food, but also constitutional disorders, as a result of which the organs of generation and those glands guarding their function receive insufficient nourishment. It is a well known fact, and long has been, that animals fail to breed when they are in a run down condition or when they are fed a deficient diet. Cary (35), quoting Hagner, states that the virility of the spermatozoa is often i n direct proportion to the general physical condition of the patient. Reynolds (34) emphasize.- the fad that it is an established principle among animal breeders that a high protein diet in both sexes is essential to full fertility. " Oligospermia with deficient vitality of the spermatozoa is not infrequently found from con- stitutional disorders. It can easily lie demonstrated in animals that both low diet and conditions of life that produce a nervous excitable state are attended by oligospermia." Animals that are closely confined, those that are over-fat (show animals), as well as those fed a deficient ration very frequently fail to breed, but exercise and change of diet soon overcome the impotency. . Dutscher, Hart, Steenbock, and other biological chemists have done extensive work to show the essential importance of vitamines and minerals in the diet. Their results indicate that animals can- not thrive and breed normally when fed a diet composed solely of the products of one plant. There must be variety, and there must be not only a correct nutritive ratio, but the mineral and vitamine content must be present as well. Cows fed on the products of one plant often failed to breed, and if conception occurred, it in- variably resulted in a premature birth, or the birth of weak and poorly nourished calves. The work of these authors is funda- mental, and brings out many important points. Is it not probable that the deficient diet results in weakened tissues which are easier prey to the invasion of bacteria .' Macnmber and Reynolds f: '.-> experimented upon white rats to determine the effect of defective diet as a cause of .sterility. They call attention to the confusion caused by the application of the term sterility to most, or all, infertile matings. They believe that failure of reproduction is. in fact, the result of decreased fertility rather than of actual sterility on the part of the two indi- viduals concerned. " There arc certainly a large number of in- tile matings which are purely functional and due to physiologic alterations or local conditions. Such physiological alterations more- over coexisl in the sterilities of pathologic origin and when un- recognized and, consequently unremedied, undoubtedly explain a 39 large proportion of the continued infertilities after operation." Is it possible for a bull to be infertile to the cows in his herd that have been fed a deficient diet, and at the same time to breed well when mated to animals outside this herd ? This is rather im- probable in practice, but there is always the possibility of its occurrence. In the experimental work, white rats were used: one strain from the Wistar Institute with a fertility of about 65 per cent, and the other from a Dr. Castle's strain with a fertility of about 90 per cent. The Wistar rats were fed in groups, each group receiving a diet deficient in a certain substance : calcium, protein, or fat soluble vitamine. To this group was added a diet deficient in both calcium and protein (war diet). These diets reduced the fertility of the groups from the original 65 per cent, to 55, 31. and 14 per cent respectively. It delayed the appearance of fertility in young rats raised on these diets, and lowered its degree in the mature animals. Most of these rats, however, though infertile to each other, bred promptly when mated to the Castle rats of known fertility. This demonstrates clearly that relative infertility of given matings does occur. One interesting feature of the work is the fact that in the matings on the single deficiency diets, four deliveries of macerated fetuses occurred and there were two more in eight deliveries from those reared on the war diet. No cases of this kind had previously occurred in this strain, which had been under observation for several years. Does this throw any light upon the cause of macerated fetuses in cattle? Micro- scopically the testes and ovaries of these infertile rats showed no observable changes, a fact which is of great importance to bear in mind. Williams, in his book on disease of the genital organs, brings out quite clearly the relation of defective diet, overfeeding, and lack of exercise, to reproductive efficiency. Xovarro (40) observed that pigeons fed on a diet without vita- mine B showed degeneration of the seminal epithelium, with hypertrophy and hyperplasia of the interstitial cells of the testis. Another author (Allen) showed that reduction in the quantity of water-soluble vitamine in the diet of rats resulted in total degenera- tion of all the germ cells, but it did not interfere with growth and development in other respects. The observations of Williams (41), in a pure bred beef herd in Hawaii, clearly demonstrate the intimate correlation between poor fodder as the result of extreme drought, and the accentuation of, or increased susceptibility to, genital infections, as demonstrated by clinical findings. The genital disorders started soon after the onset of the drought, and immediately took a downward trend with the advent of the rainy season. Judging by the work quoted, we will observe that deficient diet. though it does not always affect the general health, has a profound effect upon the genital organs of both sexes, associated with dis- turbances of spermatogenesis in the male. In most debilitated and 40 weakened conditions of the male, spermatogenesis ceases or is markedly defective. We must, undoubtedly, explain this fact upon the ground of deficient nourishment to the reproductive organs or possibly the endocrines. The vitamines have been termed nuclear nourishers, and their absence probably results in nuclear deficiency. Endocrines: Bell (42) emphasizes the fact thai not only the structure but also the function of every part of the body is in close correlation with the rest. *' This is essentially true of the ductless glands: the shadow of their influence is over all." Further he states that when we remember that the individual exists to perpetuate the species, it is not difficult to realize that the metabolic factors concerned in reproduction are the same as those related to the individual metabolism. It follows, therefore, that the ductless glands which regulate the individual metabolism concern equally the reproductive. Brown (43), discussing the same subject brings out the generalization that the sympathetic, since it is the most primitive part of the nervous system, is cl< associated with the endocrine system, a still more elemental means of communication in the body. Also since specialized reproductive cells appear before the nervous system, the organs of reproduction remain closely associated with the older chemical reactions now specialized in the endocrine glands. " The endocrine glands, the reproductive organs, and the sympathetic nervous system, there- fore, remain as a basic tripod, and it is not likely that a disturb- ance will occur for long in one limb without affecting the other two." The former author believes that the gonad keeps the other ductless alands informed of the needs of the genital tract, they in turn influencing the general metabolism. -Tump (44) sta " We are therefore justified in believing that there is a correlation of function between these (endocrine) glands and that, some cases of sterility are probably due to a derangement of this correlation." Biedl (45) concludes: " There appears to be an intimate anatomi- cal and physiological interrelationship between the different blood glands which is manifested clinically by the fact that the patho- logical disturbance of one gland is accompanied by symptoms point- ing to the functional derangement of one or more of the others. Knowing, as we do, the many sided interactivity which subsists be- tween the different internal secretory organs, it is readily con- ceivable that isolated diseases of single organs of this group are very much rarer than, at the first glance, they would appear to be. In the present state of our knowledge, the only course of investiga- tion which is open to us is to start with the known results of the functional derangemenl of any organ, and. by following these up, to seek the primary link in the pathological chain." Most workers seem to agree that a special connection exists be- tween 11k 1 normal function of the adrenal cortex and the sexual organs. Tumors of the former are usually associated with sex 41 abnormalities, and feeding young animals the gland substance seems to stimulate growth of the testes. Many arguments have been brought forward to show that the prostate produces an internal secretion. It is a well known fact that this organ atrophies after castration, and enlarges as the sex life dwindles. As has been previously stated, Serrlach and Pares reached the conclusion that the gland produces an internal secre- tion which controls the testicular functions and regulates the pro- cess of ejaculation. Also they state that if the prostate is re- moved, spermatozoa are no longer produced in the testes, and that the secretory activity of the accessory genital gland ceases. The secretion is, at any rate, a stimulus to the internal secretion of the testis. The thyroid bears a distinct biological relationship to the sexual glands. Removal of the gland results in imperfect development of the gondas, infantilism, and general torpor. Bell (42) believes that the association between the thyroid gland and the genitalia is as intimate as the relation of the pituitary to the genital functions. Of all the endocrines, perhaps the anterior lobe of the hypophysis is in most intimate correlation with reproduction. Castration results in hypertrophy of this organ, while removal of the anterior lobe usually leads to death. In those cases in which death does not ensue, it results in genital atrophy, stunting, and reduc- tion of sexual activity. In young animals, spermatogenesis ceases entirely even after partial extirpation of the anterior lobe. Biedl (45) states that " in disease of the hypophysis, derangement of sexual activity occurs very early in the course of the disease, shown in women by the cessation of menstruation, and in men by im- potence." The thymus, as is well known, is quite intimately associated with the development of the genital organs. Its normal disap- pearance is always associated with the development of sexual maturity in the individual. Hewer (46) conducted experiments to ascertain the effect of thymus feeding on the activity of the re- productive organs in the rat. She concludes in part : " Male rats appear more susceptible to the influence of thymus feeding than female rats. With moderate doses of thymus, sexual maturity in the animals treated is delayed, a phenomenon which is attributed to delayed development of the testis. With large doses of thymus, in the male, the testis is structurally affected : in the young animal in the direction of retardation of development, in the mature ani- mal in the direction of degeneration. This degeneration is con- fined to the testes. In the degenerating testis, cells of Sertoli ap- pear to be absent : the spermatogonia are present, also dividing, and may lie free in the lumen of the tubule; spermatids, many with abnormal nuclei, are shed into the lumen in large numbers; sper- matozoa are practically absent. In the later stages, only a few dividing spermatogonia appear among the debris of the other un- recognizable cells of the tubule. In the epididymis which itself is 42 normal, when the testis is showing degeneration, very few sperma- tozoa appear, in the later stages none. Many spermatids are pres- ort in various stages, and some spermatocytes. Animals in the yper-thymic eonclition appear to be sterile." The foregoing references will, I hope, serve to bring out the facts environment and diet, together with the general body meta- bolism and the endocrines, have a more or less profound effect upon the development, growth, and functioning of the genital system. In the experiments it has been shown that sterility is not necessarily accompanied by any apparent microscopical changes in the gonads, or even at times in the general body health. Nor can we exclude impotency of the male entirely even when the spermatozoa are normal in shape, and motility. Carnett, and others (38), years ago stated: " Indeed, there is abundant clini- cal proof to the effect that systemic conditions which have no ap- preciable effect upon the motility or conformation of the sperma- tozoa materially interfere with reproductive power." The entire complex genital system is inseparably linked up with the body as a whole, a fact which we must bear in mind at all times. Impediments to coitus may be due to great difference in the size the two mated individuals, psychic disturbance-, or inability to protrude the penis. Williams (17) mentions several physical impediments, as deformity of the limbs or feet, sore feet, overload- ing of the rumen, obesity, fear of falling, and paralysis. Coitus may be somewhat delayed, or even not performed as the result of severe inflammation with sensitiveness of the penis or prepuce. ' Occasionally tumors of the penis are encountered which may inter- fere with profusion of the penis, or its entrance into the vagina. Not infrequently the penis is rendered incapable of protrusion as the result of inflammatory adhesions, tuberculosis of the preputial lymph glands, etc. Excessive sexual use, within certain limits, probably has not, in itself, any material permanent effect upon the reproductive capac- ity. The frequency with which bulls used to excess break down sexually, is probably due to the devitalizing effed upon the tissues of the genital organs, this opening the way to bacterial invasion and other destructive influences. Over-use is probably not danger- unless continued over long periods, but at the same time it •s greater opportunity for infection to be introduced into the y from intercourse with large numbers of females. Llo3 r d- Jones and Hays (47) carried on very interesting experiments on the influence of excessive sexual activity of male rabbits on the perties of the semen. Their plan was to mate male rabbits in •;; succession, and study the character of the semen on the service, and every fifth service thereafter. The safe limit was twenty services in three hours. As would be expected, the volume of the semen, after the first few services, became gradually reduced in amount. " In rapidly successive services, the semen imes less viscous and tends to lose its charaetertistic milky 43 appearance until at the twentieth service, when the fluid is thin and watery. ' ' It seemed as though there was a well marked reduc- tion in the number of spermatozoa per cubic centimeter in the ad- vanced services. Successive copulations also resulted in a marked decrease in the number of motile spermatozoa, together with a shorter duration of perceptible vitality. The certainty of produc- ing impregnation at the same time became less and less. "This reduction in the per cent of effective matings when the male is sexually overworked is recognized by those engaged in animal breeding as one of the most noticeable and universal concomitants of heavy sexual service." In another paper, these same authors studied the effect of sexual excess upon the character of the offspring. In part, they conclude : "By no means thus far used has any inferiority of progeny from the heavy sexual service been discovered. They are fully equal if not superior to progeny from very light service of male." Infection is without doubt the greatest single factor capable of producing functional and anatomic changes resulting in varying degrees of impotency and sterility. The changes produced range from the addition of the toxic products of bacterial growth to the seminal fluid, to the complete destruction of the parenchymatous tissue of one or more of the contributing sexual glands. Anatomic changes are by no means essential to the production of lowered fertility. As has been previously stated, the work on veal calves indicates that the genital organs of young bulls are normally free from bacteria. Likewise in 'normal adult animals, the bacterial content of the genital organs is as a rule low or negative. It is possible that a certain flora is normal for the tract at sexual maturity, as in several other organs of the body, but under the strain of sexual excess, defective diet, or other weakening in- fluences, these organisms may become pathogenic. Streptococci and staphylococci have at times been found in apparently normal parts of the body, and at other times they are found associated with severe pathological lesions in the genital tract. The degree of pathogenicity is of course difficult to determine, except as we find the bacteria associated with abnormal conditions. Carpenter (9), however, injected streptococci into the genital tracts of female calves and produced lesions resembling very closely those from which the organisms had been isolated in adult sterile animals. Per- sonally, I am inclined to believe that the genital organs normally are free from bacteria, or if any are there they are better able to multiply under the strain of devitalization of the tissues. Bacterial invasion, however, does take place quite frequently, but the paths of entrance of the -organisms are somewhat proble- matical. Hematogenous origin is always possible, though it is rather difficult to definitely implicate this mode of entrance. The urethra is perhaps the easiest and most common path for the entrance of bacteria, though even here it is not possible to make definite assertions. Contiguous spread of infection from neigh- •II boring structures is very probable in some cases, particularly in pelvic peritonitis. The bacteriological results hardly bear out the theory of Williams thai the organisms lie dormant in the genitalia of the animals until the advent of sexual maturity, at which time they acquire pathogenic powers. < >n the other hand, his clinical observations seem to indicate that this may be possible. Calves suffering from "calf infections" frequently do harbor organisms in their genital organs, but whether or not they persist there till sexual maturity is a matter of conjecture. The most logical theory seems to be that animals from herds in which genital infections are very severe, or those that have had severe attacks of scours or pneumonia, are more susceptible to those infections, due to the early lowering of their vitality. One bull in the department herd certainly had a severe ordeal as a calf, but as a mature bull he was highly fertile. Moderate sexual use and proper sexual hygiene probably had much to do with this. In the bull, infection of some part of the genitals, during some period of life, is very con- stant, however, whether or not it is productive of observable changes in his breeding efficiency. The finding of the fine connec- tive tissue strands and tufts on the serous surface of the tail of the epididymis of practically all bulls examined, both sterile and fertile, indicates past or present infection of the scrota] sac. The vesicles and tail of the epididymis are, as stated previously, the most commonly invaded, tissues of the tract. The testes are frequently involved. While it is difficult to obtain irreproachable proof that the bull is a disseminator of genital infections, the findings of clinici quite clearly indicate that this is true, and laboratory methods tend to support tins assumption. Williams believes that not only may the bull infect the female with organisms which interfere with the given conception, but that he often implants there organ- Avhich interfere with future pregnancies, and even with the life of the individual in some cases. The high abortion and ster- ility rate following the use of certain sires, and the appearance of characteristic infections after service to certain bulls, clearly in- dicate that in all probability the bull does eliminate with his semen those organisms winch produce lesions in his genital organs, and are capable of infecting the female. \Y. L. Williams (48) cites the case of a pure bred herd in which breeding had pro- gressed satisfactorily until heifers had grown to breeding age and cond bull was obtained. " Some cows of the old herd were assigned to the young bull which had not previously been in service. The cows bred to the old herd bull continued to breed normally. The cows and heifers bred to the new bull conceived with difficulty ov not at all. Those which conceived mostly ted, and those which calved had metritis and retained fetal membranes. The two first cows in which pregnancy terminated died of metritis." 1 have frequently had semen samples sent in from bulls thai were no1 only failing to gel cows with calf, but 45 following each service the females showed a severe vaginitis. W. W. Williams worked in a herd in which service to certain bulls was in each case followed by a severe vaginitis and cervicitis, only to be followed later by a characteristic salpingitis. Vaginal smears taken before and after service, in many in- stances, show that in all probability bacteria, especially the strep- tococci, were deposited there with the semen. These results have been obtained upon several occasions, at which time the vagina was usually douched prior to each service with sterile saline solu- tion. Streptococci and other organisms have been isolated from the vaginal samples obtained by this method. In most cases, they were absent from samples taken before service. Extraneous con- tamination, and error, must be taken into consideration, but the results tend to bear out clinical observations that the bull is prob- ably a disseminator of some infections associated with the genital organs of both sexes. At any rate, organisms have been isolated repeatedly from the genital organs of the bull, of the same bio- logical character as those which are associated with sterility, abor- tion, and allied phenomena in the female. In the absence of obstruction in any part of the tract, there is nothing to hinder infection from gaining access to the seminal fluid, and being ex- creted during ejaculation. Hopper (18) states: "A diseased bull may manifest non-fertility or decreased potency in different ways — by repeated service to apparently normal females without conception, by a high abortion rate in females that have been apparently normal, by characteristic infections following the use of any particular sire, or by abnor- malities in the breeding tract noted by rectal or physical pal- pation." Admittedly, Bad. abortum has little affinity for the genitalia of the bull, though Schroeder states that the bull harbors the organisms in -his seminal vesicles and that they are eliminated with the semen. Other authors have occasionally isolated the or- ganism from the vesicles, testes, or both. Schroeder 's theory that infection of the female occurs indirectly by contamination of the fodder with the semen is probably rare in occurrence. The very limited number of cases in which investigators have demonstrated the presence of the organism in the male genital organs, and the apparent immunity of the bull to the bacterium as determined by the agglutination reaction, seem to indicate that he plays a small part in the spread of this type of infection in the herd. On the other hand, it would seem that he is at times intimately associated with the spread of certain other organisms that interfere seriously with herd reproductivity. The diagnosis of infertility and sterility rests upon a thorough physical examination of the genital organs, together with a de- tailed study of the semen. The history of the animal and herd involved must also be very carefully inquired into, especially the part covering the result of every service by the sire in question. 46 Besides this, we must always consider all factors which have a bearing upon the subject, remembering the physiology of each part, and the role it plays in reproduction. Bacteria gain entrance to many parts of the tract, where they multiply and probably add toxic products to the seminal fluid, altering its biological char- acter and resulting in partial or total destruction of the secretory tissues. The testes, epidid.vmes, vesicles, prostate, and Other parts. each contribute their essential pari to the semen, abnormalitie any one of which, as a rule, result in interferences with reproduc- tion. If the vesicles are involved, we must bear in mind just what is the part played by their secretion, and what is the probable result if their essential elements are not added to the semen. Jn like manner, we must consider the prostate, whose secretion stimu- lates the vitality of the sperms, and adds fluid bulk to the semen. Extirpation of the vesicles or prostate alone results in lowered fertility, without altering the sexual desire, while removing both glands produces total sterility. Partial or total destruction of the parenchyma of either gland produces the same effect as extirpa- tion, in that its function is altered or entirely absent. The semen should be examined, not only for the number of spermatozoa and the percentage of those that are motile, but for the duration and type of motion. Normal semen, when first ex- amined under the microscope, shows a field closely packed with highly motile spermatozoa. In every study of the semen, however, we must bear in mind the temperature and other conditions under which it has been kept since emission. On the other hand, semen from hulls of lowered fertility shows changes ranging from mild disturbances such as sluggish motility and a slight decrease in the number of sperms present, to aspermia, or total lack of motion. Normal semen, when compared with abnormal specimens, as a rule presents distinct differences, either in motility, staining properties, or structure of the spermatozoa. Impotent bulls, however, may show at times few or no observable changes in their genital organs. The only assumption her* 1 is that the condition probably is of endocrine origin, or is some functional disturbance. Of oligo- spermia Keynolds states: "Oligospermia, with normal motility and vitality, is not absolute sterility, but is of high importance because the percentage of destruction of spermatozoa during their passage through the genital canal of the female is so enormous thai the possibility of impregnation by semen which starts out with a delicieiit number is always poor. When the genitals of the female partner are in a condition which is even moderately hostile to the spermatozoa, impregnation by such semen becomes so unlikely as to be noi even a probability." Motility may be lacking in a small number, its absence may be observed in a large percentage, or even in all those in the field. as in necrospermia. On the other hand, the motility may be sluggish or of abnormal types in variable percentages. Sperms 47 with sluggish motility are always low in vitality, and have weak powers of insemination, as the motion lasts but a comparatively short time. The vitality may be but moderately lowered, so that although the sperms are highly motile when ejaculated they soon lose their power of propulsion. The type of motion is likewise an indicator of lack of vitality. The "progressive vibratile" motion described by Reynolds should proceed to a high degree for a long period before the "undulatory tactile" or bunting types of motion appear. Early appearance of these two latter types indicates in most cases a marked lack of vitality of the elements. The motion should be vigorous and lasting, for, as stated by Reynolds, "noth- ing is more certain than that spermatozoa of merely moderate vitality seldom impregnate a female." The early precipitation out of the "Boettcherchen" crystals is very characteristic of oligospermia, and impotent semen. The theory here is that crystals do not precipitate out when a fluid is actively moving, but soon do so when the fluid is motionless. The sediment which normally makes up about two-thirds of the sample is usually decreased in abnormal samples. The semen itself should be observed for unusual viscosity or a thin watery condition. (Totting should occur readily after emission, but the clot soon liquefies to some extent, allowing the spermatozoa to become more active. This clotting is, of course, to protect the delicate sperms from the hostile secretions of the vagina. The presence of immature and deformed types of sperms rep- resents some disturbance of spermatogenesis, but it is difficult to explain the significance of these forms. They are seldom seen in normal samples, and undoubtedly none are capable of producing impregnation. In the case of minor abnormalities of stajning re- actions, the sperms are probably deficient in nuclear material or otherwise altered so that probably they are incapable of reaching and uniting with the ovum. When impregnation does occur in these cases, weak offspring undoubtedly result in many instances. The work has by no means progressed to the point where one may, by an examination of the semen, determine the degree of impotency with great accuracy, or even whether the animal may be restored to sexual health by proper hygienic and therapeutic treatment. Relatively, the greater the changes in the semen and spermatozoa, the less the chances of impregnation. Infertility to any marked degree, is, however, usually accompanied by cor- responding changes in the seminal fluid and its germinal elements. Examination of the semen is, and probably always will be. simply an aid in reaching a diagnosis. While abnormalities of the semen and spermatozoa are associated with sterility or in- fertility, it is unwise to lay too much emphasis upon this method of diagnosis alone, especially with regard to the making of a definite prognosis. When large numbers of abnormal spermatozoa are present in the semen, we are safe in saying that the animal 48 is. at the time, of lowered degree of fertility. One should be very cautious, however, in foretelling how long the condition will last, or if the animal may in time be restored to full fertility. Ster- ility, due to organic disturbances, probably seldom yields to treat- ment, but when it is due to functional disorders resulting from defective diet or lack of exercise, the condition is frequently reme- died by overcoming the cause. Lack of exercise and overfeeding seem to be etiological factors in a fair percentage of cases. Besides abnormalities of the male genital tract, we must always consider the numerous factors in the female that may kill or weaken the sperms. Impediments to successful coitus may be present in the form of vaginal constrictions, abnormally short or small vagina, or other deformities. Hostile exudates, mechanical obstructions, and other factors may interrupt the progress of the sperms at any point in the tract. Although little is known definitely regarding disorders of the endocrines in the hull and their relation to reproduction, the work in human medicine and experimental researches upon laboratory animals warrant thoughtful consideration of these factors which are by no means insignificant. In the future, these glands will no doubt receive more and more attention in their relation to the genital organs and reproduction. Conclusions 1. The genital organs of the bull quite frequently undergo pathological changes, due to infection with the same varieties of microorganisms associated with genital infections in the female. 2. In all probability, these microorganisms are frequently elimi- nated with the semen and infect the female during copulation. 3. Past or present infection in the genital organs of all the bulls so far examined was evidenced by the presence of the fine connective tissue tufts and strands upon the tunica vaginalis, par- ticularly that part covering the tail of the epididymis. 4. Lowered sexual capacity is. as a rule, accompanied by demonstrable changes in the semen. 5. A study of impotency and sterility includes not only a thor- ough study of the genital organs, but also those extrinsic factors which govern reproduction either directly or indirectly. 6. A thorough knowledge of the anatomy and physiology of the male genital organs is fundamental to a clear understanding of the problem. I am much indebted to Drs. W. 1.. and W. W. Williams for some of the material, and for helpful co-operation in the early part of the work; to Drs. ('. M. Carpenter and R. K. Birch for many helpful suggestions; and to Dr. J. N. FroM and others who so kindly co-operated by placing at my di>- posal samples of semen and sonic of the genital tracts. 49 Bacteriology of the Genital Tracts of Normal Young Calves Group I Number Right sem. vesicle Left sem. vesicle Right testis Left testis 1 3 4 5 Staph, albus. 6 7 9 Staph, albus. Staph, albus. 12 13 14 50 r. 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Die Aetiologie des seuclienhaften ( infectiosen ) Verwerfens. Zeit Med. Band I, S. 241, 1897. Das seuclienhaften Yerwerfen der Binder. Arch. wiss. u. prakt. Thierheilk., Band 33, S. 312, 1007. 2. Law, James. Contagious Abortion of Cows. Circular No. .3. Agr. Experiment Sta. University of California, June. 1903. 3. Report of English Commission of Epizootic Abortion, Appendix to Part I, p. 17, 1909. 4. Hadley, F. B., and Lothe, H. The Bull as a Disseminator of Contagious Abortion. Jour. Amer. Vet. Med. Assoc, L, 1916-17, p. 143. 5. Hadley, F. B. Contagious Abortion Questions Answered. Bulletin Xo. 296, Oct. 1921, Agr. Experiment Sta., University of Wisconsin. 6. Buck, J. M., Creech, G. T., and Ladson, H. H. Bacterium Abortus Infec- tion of Bulls (Preliminary Report). Jour. Agr. Research, August, 1919. 7. Schroeder, E. C, and Cottox, \Y. E. The Bull as a Factor in Abortion Disease. 8. Cotton, W. E. Proceedings of A. V. M. A., p. 851, 1913. 9. Carpenter, C. M. Report of the New York State Veterinary College, Cornell University, 1920-21. 10. Rettger, L. F., and White, G. C. Infectious Abortion in Cattle. Storrs Agr. Exp. Sta. Bulletin No. 93. January, 1916. 11. McFadyean, Sheather and Minett. Researches Regarding Epizootic Abortion. Jour, of Comp. Path, and Therap., XXVI. 142, 1913. 12. Schroeder, E. C. Bureau of Animal Industry Investigations of Bovine Infections Abortion. Jour. Amer. Vet. Med. Assoc, LX, p. 542. February, 1922. 13. Barney, J. D. Recent Studies on the Pathology of the Seminal Vesicles. Bost. Med. and Surg. Jour.. CLXXI. 1914, 59. 14. Williams, W. L. The Diseases of Bulls. Cornell Veterinarian. X, 94, April, 1920. 15. Williams, W. L. Report of the Xew York State Veterinarv College, Cornell University, 1920-21. 16. Williams, W. W. Technique of Collecting Semen for Laboratory Examination with Review of Several Diseased Bulls. Cornell Vet. X, 87, April, 1920. 17. Williams, W. W. Diseases of the Bull Interfering With Reproduction. Jour, of Amer. Vet. Med. Assoc, LVIII, 29, October, 1920. 18. Hopper, E. B. Herd Efficiencv from the Standpoint of the Veterinarian. North Amer. Vet., Ill, 71, February, 1922. 19. Williams, W. W. Observations on Reproduction in a Purebred Dairy Herd. Cornell Veterinarian, XII, 19, January, 1922. 20. Walker, K. M. The Diagnosis and Treatment of Sterility in the Male. Lancet, CCI, 228. July 30, 1921. 21. Pende. Endocrinoglia-Pathologia E Clinica. Abstract in Endocrinologv, II, 42. 22. Ferguson, J. S. Normal Histology and Microscopical Anatomy. 23. Ellenberger. Vergl. mikroskop. Anatomie der Haustiere. II. 24. Courrier, M. R. On the Existence of a Secretion of the Epididymis of the Hibernating Bat and Its Significance. C. R. Soc de Biol. (Paris), 1920. LXXXIII, 67-69. 23. Stigler, R. Abstract in Jour. Phys. et Path. Gen. (2) CLXXI, 273. September, 1918. 243. Disselhorst. Ansfuhrapparat und Anhangsdrusen der mannlichen Geschlechtsorgane. Oppel's Lehrbuch der vergleichenden mikr. Anat. der Wirbeltiere, IV, Jena, 1904. 27. Rytina, A. A. The Verumontauum. With Special Reference to the Sinus Pocularis: Its Histology, Anatomy, and Phvsiologv. Jour. Urology. I, 1917, 231. 28. Fish, P. A. The Spermatic Secretion and Its Ultraparticles. Cornell Veterinarian, October, 1921. 60 29. Marshall, F. II. A. The Physiology of Reproduction. 30. Kingsbury, B. F. Professor of Histology and Embryology, Cornell Uni versity, Ithaca. New York. 31. Walker. Arch. f-Anat. u. Entwicklungsgesch., 1899, und Arch. f. Anat. u. Physiol. 1899. 32. Boettcheb, W. On the Significance of the Secretions of the Male Acces- sory Genital Organs. Munch, med. Wchnschr. 1920. 07, 1, p. 44. 33. Editorial. Jour, of Amor. .Me.!. Asm,,.. LXXVII, No. i. 42. July 2. 1921. 34. Reynolds, E. Fertility and Sterility. .lour, of Amer. Med. Assoc., I. Mil, I L93 HH'.i, October 21. L916. 35. Cart, \V. II. Examination of Semen With Special Reference to Its Gynecological Aspects. Amer. Jour, of Obstetrics and Disea Women and Children. LXXIV, No. 4. L916. :iii. WOLF, C. G. L. The Survival of Motility in Mammalian Spermatozoa. Jour, of Physiol. IV. 246, August 3. 192-1. J7. BROWN, J. H. The Use of Blood Agar for the Study of Streptococci. Monograph of the Rockefeller Institute for Med. Res., 1919, IX. 38. GARNETT, J. B., and others. The Surgical Treatment of Sterility due to Obstruction at the Epididymis Together with a Study of the Morphology of Human Spermatozoa. Univ. of Penn. Med. Bulletin, March. 1902. 39. Reynolds. E. and Macomuer, D. Defective Diet as a Cause of Sterility. Jour, of Amer. Med. Assoc, LXXVII, Kill. July lfi, 1921. 40. Novarro, P. A. Tissues of the Testicle and Antaminosis. Gazetta degii Ospedali (Milano), 1920, XLI, 424. 41. Williams, W. L. Observations on Reproduction in a Pure Bred Beef Herd. Cornell Veterinarian. January, 1922. 42. Bell, W. B. Correlation of Function: With Special Reference to the Organs of Internal Secretion and the Reproductive Svstem. Brit. Med. Jour., 1920, 1, 787. 43. Brown, W. L. The Principles of Internal Secretion. Brit. Med. Jour. 1920, IT, 687-691. 44. Jimp. Discussion in Penn. Med. Jour., XXV, 81. 45. Biedl. A. The Internal Secretory Organs. 46. Hewer. E. E. The Effect of Thymus Feeding on the Activity of the Reproductive Organs of the Rat. Jour, of Phvsiology. XLVII, 1913- 1914, 479. 47. Lloyd-Jones, 0. and Hays, F. A. The Influence of Excessive Sexual Activity of Male Rabbits. 1. On the Properties of the Seminal Dis- charge.' Jour, of Expcr. Zoology, 1918, XXV, 463. 4S. Williams. W. L. Improvement of the Reproductive Efficiency of Cattle. North Amer. Veterinarian, HI. May. 1922. Description of Plates Plate I. Fig. 1. Diagrammatic sketches showing the development of a spermato- zoon from a spermatogonium. Fig. 2. Diagrammatic sketch showing the minute structure of a sper- matozoon. The middle pieee i~ made comparatively thick in order to bring out the finer structures. Adapted from Ellenberger. Pla ii: II. Fig. 3. Testicle of bull, showing extensive degeneration and necrosis. Plate III. Fig. 4. Tuner part of wall of ductus deferens. Normal, x 230. Fig. ■">. Same, but showing extensive degeneration and exfoliation of the lining membrane, x 230. Plate IV. Fig. 6. Ductus deferens, showing entire exfoliation of the lining mem- brane. The lumen i- filled with a cellular debris, x 50. 61 Fig. 7. Same, showing the degeneration of the lining membrane, and debris in lumen, x 230. Plate V. Fig. S. Seminal vesicle of bull. High power section showing the normal structure of the vesicular cavities. Fig. 9. Seminal vesicle of bull. Low power. The membrane lining the cavities is degenerated and exfoliated. The cavities are filled with cellular debris, and exudates. There is some increase in the interstitial tissue, and atrophy of some of vesicular cavities. PXATE VI. Fig. 10. Low power section of seminal vesicle of bull. The condition is about the same as in Fig. 9 except that it is not quite as severe. Fig. 11. Same as Fig. 10. High power. Plate VII. Fig. 12. Testicular tubule showing normal spermatogenesis, x 230. Fig. 113. Testicular tubule showing no evidence of mitosis, x 230. Fig. 14. Testicular tubule. The spermatogenic epithelium is beginning to degenerate and become cast off into the lumen, x 230. , Fig. 15. Testicular tubule, showing almost total exfoliation of the spermatogenic epithelium, x 230. Plate VIII. Fig. 16. Testicular tubule. The seminal epithelium is entirely degen- erated. The membrana propria is markedlv thickened. x 230. Fig. 17. Testicular tubule. The tubule is undergoing atrophy and degeneration. The interstitial connective tissue is much increased in amount, x 230. Fig. 18. Same as Fig. 17, except that it is in the more advanced stages. Fig. 19. Testicular tubule. The spermatogenic epithelium has under- gone a sort of hydropic degeneration. The interstitial con- nective tissue has become much increased in amount and has undergone cellular infiltration. Plate IX. Fig. 20. Normal structure of epididymis tubule, x 140. Fig. 21. Epididymis tubule showing exfoliation of the lining membrane, and cellular debris in the lumen, x 140. Fig. 22. Atrophy and degeneration of epididymis tubule. The inter- stitial connective tissue is much increased in amount, x 140. Fig. 23. Same, but in more advanced stage, x 140. Plate X. Fig. 24. About the same as Fig. 22. x 140. Fig. 25. Marked degeneration of epididymis tubule. There is a cellular infiltration of the interstitial tissue, x 60. Plate XL Fig. 26. Spermatozoon, showing constriction at middle of head. The head is also somewhat contorted, x 670. Fig. 27. Spermatozoon. Pear shaped head, x 670. Fig. 28. Spermatozoon. The head is quite long, and pointed at its posterior end. x 670. Fig. 29. Spermatozoon, showing a constriction at middle of the head, x 670. Plate XII. Fig. 30. Microcephalic spermatozoon, x 670. Fig. 31. Spermatozoon. The head is small, and pear shaped, x 670. Fig. 32. Macrocephalic sperm. The middle piece is much thickened. x 070. Fig. 33. Tailless spermatozoa, x 670. /Vac/cu.S Cen/rosomes Ce-ntrospkere CoTmec~t/r>a f?ece of tcuiL of fro it Qcrosorne Disc ardedCu top I asm need Liaht Inner hodu Cntbr/or J-'odule. A/ecA Posterior nodule Spiral filament Cutoyb/asrn/c sheath. Cmiular nodutc S77ooluc.ru.yn. PLATE II Fig. 3. PLATE III Fig. 4. Fisr. 5. PL VI E IV Fig. G. f ~ jt-%. 1