A study of some factors influencing fertility and sterility in the bull

by Herbert Lester Gilman

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 microscopical 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 number 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 following 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.

Testes: The testes seldom presented gross alterations of structure except for abscess formation, which, according to Williams, occurs more frequently in the bull than in any of the other domesticated animals. He also states that 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 with a history revealing 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 at 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 uncertain 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 centimeters. The tunica albuginea presented a thickness of six millimeters, and was made up of firm sclerotic tissue. The epididymis 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 Bact. abortum.

Microscopically, changes are quite common and varied in character. 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 case in the left 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. Likewise, 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 Fig. 15. 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 gland, no sperms are formed. Not infrequently 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 6. The cells are several layers deep as in the normal condition, but they are not dividing. This condition is shown in Fig. 13.

In the more chronic forms, the tubules become atrophied, and frequently disappear entirely. The membrana propria may become thickened, due to excessive connective tissue formation, or infiltration with serum or exudate. On the other hand, a distinct atrophy may occur. The stroma of the organ not infrequently 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 inflammations, the entire organ becomes enlarged, markedly hyperaemic, and infiltrated with leucocytes. Necrotic areas appear here and there in the parenchyma. The rete often shows a marked degeneration of the lining epithelium, and atrophy caused by increase of the interstitial connective tissue.

Epididymis: This organ not infrequently presents gross abnormalities, and very often is pathological on microscopic examination. 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 tissue. In adult bulls it is exceptional not to find at least slight evidence of some previous inflammation. In all of the numerous bulls examined, 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 hyperaemia, 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 degenerated, 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 pyogenic types of inflammation.

Ductus Deferens: 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 infrequently show a mild type of degeneration and exfoliation, or in the more chronic forms, the entire membrane degenerates and disappears. In man, the duct occasionally becomes occluded, but so far I have failed to find this condition in the bull. Undoubtedly, when the occlusion does 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 the 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. Occasionally the abscesses rupture and discharge their contents into the rectum. Dr. Williams 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. On 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 distinct 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 condition 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 superficial thickening, or it may extend in between the lobules.

4. The Peri-vesicular or “pan-inflammatory” Type usually is the result of severe inflammation of the vesicles, with probable rupture 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 encountered, 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 inflammation 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 interstitial tissue is in no way affected, but at times it is thickened by oedematous exudates, leucocytes, and fibrin. The chronic interstitial form is characterized by a considerable increase of connective tissue,—producing marked atrophy, or even complete obliteration of the glandular cavities. Microscopically the suppurative form may be diffuse over the entire gland, or as stated previously, may be in the form of localized abscesses, with or without 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 undoubtedly 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 subjected 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 Cowper’s glands, as well, occasionally undergo inflammatory changes.

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 internal 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. With this in view, we must remember that disease of any of the contributing organs is a potential 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. This 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, hypofunction 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 protecting 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 semen was quite thick, of a yellowish green color, and of a distinctly acid reaction. The secretion from the vesicles was later found to be of this same character, and was due to a Ps. pyocyaneus infection. The vesicles were highly inflamed and degenerated. The spermatozoa were in this case markedly decreased in number, and devoid of motility.

The early precipitation of the “Boettcherchen” crystals seems to be intimately connected with sterile semen, or spermatozoa of lowered vitality. Likewise, a decrease in solid matter is often seen in a deficient secretion. In 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.

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 swimming,” 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 recognize, 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 lower 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 obtained 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, varying 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 progressive movement should, under proper conditions, survive for a considerable time before it gives way to the second, and slower type. The cells frequently early bunt 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 entirely. I have seen one specimen in which the sperms all tended to clump. Whether this was the result of some agglutinative substance in the vaginal secretion is problematical. I have seen several specimens of semen in which practically all the sperms were motile when first 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 ejaculated into the vagina, large numbers of them are destroyed or become 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 diminished in proportion to the number of dead or defective sperms.

Aspermia: Absence of spermatozoa in the semen is rarely encountered, and is probably due either to total cessation of spermatogenesis, 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 development 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, but a clearer understanding of them can best be obtained by a review of the process of spermatogenesis.

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. Some of these intermediate types are large oval cells without distinct nuclei and as a rule with poorly developed tails. Cells with no tails or distinct nuclei, those with protoplasmic appendages to the head or tail, and various other types, are occasionally encountered. Most of these are motionless and incapable of producing 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 supply 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 represent a degenerative process induced possibly by abnormalities 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 appendages, 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 development, 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 constriction 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 uniform thickening, or as a bulging appendage. Rudimentary development of the tail, the presence of two poorly formed tails, and defective development of the connecting piece occur rather infrequently.

All these immature and defective types are, as a rule, motionless, 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 they are absent. They probably are not the result of the methods used in fixing and staining. Some think they are slightly immature types, or that the condition is produced by abnormal contractions of the tail. The majority of sperms, however, especially those from highly fertile bulls, do not show this type at all.

The most common changes in the spermatozoa, are those in which there is a separation of the head from the tail, and degeneration of 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. The separation, in the majority of cases, indicates some lowering of 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 are very common. The cell membrane, which is normally distinct and sharp, becomes blurred in outline. Normally, the head takes a good differential stain, the anterior part staining lightly, and the posterior part somewhat deeper. The nucleus is distinct in outline and well defined. The lighter “inner body” stands out in well stained specimens. As the result of degeneration, the whole head may take the stain uniformly, either slightly or much deeper than normal, according to the degree of degeneration. The whole problem of staining, however, depends very much upon the methods used, and the care 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.