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B. CRABO and B. GUSTAFSSON

Summary.

Sodium and potassium were measured in epididymal plasma at six different levels of the epididymal duct in individual bulls, and the sperm concentration was estimated at the same levels. The sperm concentration was very low in the rete testis and the proximal part of the caput, but increased continuously to a maximum at the proximal end of the corpus. It fluctuated thereafter, being lower in the distal corpus and slightly higher in the cauda. In the rete testis the sodium content of the plasma was high and the potassium content low. The concentration of sodium decreased in the caput and proximal part of the corpus, where the potassium concentration reached its maximum, thus indicating a selective resorption of sodium with the fluid. In the rest of the corpus the concentration of sodium and potassium remained unchanged but in the cauda the concentration of both decreased markedly.

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B. CRABO and L.-E. APPELGREN

Alpha-chlorohydrin (3-chlor-1,2 propanediol) possesses antifertility properties when administered to male rats, hamsters, guinea-pigs, rams and monkeys (Ericsson, 1968; Ericsson & Norland, 1970). The compound is thought to exert its action on the epididymis, since high doses of α-chlorohydrin cause vascular disturbances in the proximal part of the epididymis (Gunn, Gould & Anderson, 1969; Samojlik & Chang, 1970; Kreider & Dutt, 1971). There is also a time delay between administration of the drug and the onset of inhibition of fertility (Ericsson, 1968; Turner, 1971). Impaired sperm motility which has been observed (Coppola, 1969; Samojlik & Chang, 1970), has been thought to be due to oxygen deficiency in the epididymis following vascular trauma. Coppola (1969) and Gunn et al. (1969) suggested that α-chlorohydrin might act as a metabolic antagonist to

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B. CRABO, B. GUSTAFSSON, L. NICANDER and A. R. RAO

Sperm counts from different levels of the male ducts have suggested a selective removal of abnormal spermatozoa in the epididymis (Orgebin-Crist, 1964; LeRoy-Gilette, 1966; Roussel, Stallcup & Austin, 1967). Phagocytosis of spermatozoa by epithelial cells, however, has rarely been reported (Nicander, 1963) except in cases of epididymal obstruction (cf. Orgebin-Crist, 1969). The present report on frequent phagocytosis of abnormal spermatozoa in the efferent ductules of a bull adds material to the discussion on sperm elimination in the male genital ducts.

The bull was of the Swedish Red and White Breed, 15 months old and purchased as normal. It had served a few times and produced offspring. During 2 months' stay at the clinic, its semen picture was within the normal limits given by Lagerlöf (1934),

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M. S. Kosco, K. J. Loseth and B. G. Crabo

Summary. Development of the prepubertal seminiferous tubules of the right testis was characterized morphometrically every 14 days from 10 to 122 days of age in intact boars (I) and boars hemicastrated (HC) on Day 10 of life from two herds (Trial 1 and Trial 2). Comparisons were made between the remaining testis of Group HC boars and one testis in Group I boars. By 38 days of age seminiferous tubule length in Group HC boars was double (P < 0·0001) that in Group I boars. Seminiferous tubule length did not differ between trials within treatments. The diameter of the seminiferous tubule was similar in Group HC and I boars but was greater (P < 0·05) in Trial-1 than Trial-2 boars from Day 80 to 122 of life. Relative mass (mass of tissue/body mass) of Sertoli cells became 2-fold greater (P < 0·0001), in Group HC than in one testis of Group I boars by 38 days of age and this difference was maintained throughout the experimental period. The relative mass of Sertoli cells was greater (P < 0·05) in Trial-1 than Trial-2 boars within each treatment between 80 and 122 days of age. The relative mass of gonocytes was similar for all groups and treatments of boars. By 122 days of age the relative mass of spermatogenic cells was greater (P < 0·05) in Group HC than in one testis of Group I boars and greater (P < 0·01) in Trial-1 than Trial-2 boars within each treatment. Onset of spermatogenesis was first observed at 80 and 94 days of age in boars in Groups HC and I, respectively. Development of seminiferous tubule lumen was first observed at 94 and 108 days of age in boars in Groups HC and I respectively. Seminiferous tubule lumen, taken as a measure of fluid secretion of the Sertoli cells, occupied a greater (P < 0·01) portion of seminiferous tubule in Trial-1 than Trial-2 boars within each treatment at the end of the experimental period. It is concluded that neonatal hemicastration of boars rapidly caused a compensatory seminiferous tubule elongation apparently due to Sertoli cell proliferation and an earlier onset of spermatogenesis. However, the gonocytes do not proliferate until they transform into spermatogonia.

Keywords: boar; testis; hemicastration; seminiferous tubule; spermatogenesis

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K. J. Rozeboom, M. H. T. Troedsson and B. G. Crabo

The objective of this study was to characterize the uterine leukocyte influx after artificial insemination (AI). After detection of oestrus with a boar at intervals of 1.5 h, seventy-two gilts were randomly assigned to a 2×3×4 factorial arrangement. AI was performed with 100 ml extended semen containing 5 × 109 spermatozoa (semen; n = 36) or 100 ml VSP semen extender (extender; n = 36) at one of three times after detection of oestrus: 12, 24 or 36 h (n = 24/time). The uterus was lavaged at 6, 12, 18 or 24 h (n = 18/time) after AI to determine the total number of uterine leukocytes. In addition, uterine lavage was performed on nine untreated gilts immediately after the detection of oestrus to establish a baseline number of leukocytes. The leukocyte response in all samples consisted predominately (92–99%) of polymorphonuclear neutrophilic granulocytes (PMNs). The mean number of PMNs recovered from the uteri of gilts treated with semen was greater than in gilts treated with extender and in untreated gilts (P < 0.01). The greatest number of PMNs in semen-treated gilts was found 12 h after AI (P < 0.01), and this number was sustained for 24 h. In contrast, the number of uterine PMNs recovered from extender-treated gilts reached a peak at 6 h and had declined by 12 h after AI (P < 0.05). It was concluded that an extensive influx of PMNs into the uterus is a normal sequence to AI. The consequences and importance of semen-induced uterine leukocytosis needs further investigation.

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R. E. BOWER Jr, E. F. GRAHAM and B. CRABO

Efforts to study the physiology and chemistry of epididymal spermatozoa have been hampered by the limitations placed on the investigator by the source of his materials. Earlier studies (Mann, 1959, 1964; Crabo, 1965) have relied on epididymides obtained at castration or following destruction of the experimental animal, procedures which severely limit the amount of data obtained from one boar. Attempts to cannulate and expose the vasa deferentia of the boar (Wierzbowski & Wierzchos, 1969; Johnson, Pursel & Kraeling, 1971; Einarsson, 1971) have had limited success because of exudative contamination, changes in sperm morphology and difficulty in maintaining a patent duct. Ninety-three days has been the maximum amount of time that epididymal spermatozoa and fluids have been collected by this method (Wierzbowski & Wierzchos, 1969).

Partial blockage of the seminal secretions of the accessory sex glands can be accomplished by injection of large doses of atropine, a parasympathetic-blocking agent (Dziuk, 1959;

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M. S. Kosco, K. J. Loseth and B. G. Crabo

Summary. Development of the prepubertal interstitium of the right testes was characterized every 14 days from 10 to 122 days of age in intact boars (I) and boars hemicastrated (HC) at 10 days of age from two herds (Trial 1 and Trial 2). Comparisons were made between the remaining testis of Group-HC boars and one testis in Group-I boars. The relative mass (mass of component/body mass) of interstitium was 151% greater (P < 0·001) in Group-HC than Group-I boars by 52 days of age. The relative mass of interstitium was greater (P < 0·01) in Trial-1 than Trial-2 boars within each treatment from 80 to 122 days of age. The relative mass of interstitial space was 76% greater (P < 0·05) in Group-HC than in one testis of Group-I boars by 52 days of age and greater (P < 0·05) in Trial-1 than Trial-2 boars within each treatment from 80 to 122 days of age. The relative mass of Leydig cells was 254% greater (P < 0·0001) in Group-HC than Group-I boars by 52 days of age and remained greater (P < 0·05) in Group-HC than Group-I boars from 52 to 122 days of age. By 52 days of age the relative mass of Leydig cell nuclei and cytoplasm was 235% and 265% greater (P < 0·0001) in Group-HC than Group-I boars, respectively, and both remained greater (P < 0·05) in Group-HC than in Group-I boars until 122 days of age. The relative mass of small vessels was 86% greater (P < 0·01) in Group-HC than Group-I boars from 24 to 66 days of age and was similar in Group-I and Group-HC boars from 66 to 122 days of age. The relative mass of Leydig cells, nuclei and cytoplasm as well as small vessels was similar between trials of boars within each treatment. Neonatal hemicastration of boars at 10 days of age therefore resulted in an overcompensation in numbers of Leydig cells, measured as nuclear mass, whereas the increase in vascular development and interstitial space did not fully compensate the loss of testicular tissue. The cytoplasm to nuclear ratio reflected the steroid production of the Leydig cells which was related to pubertal tubular development rather than treatment.

Keywords: boar; testis; hemicastration; interstitium; Leydig cell

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B. CRABO, B. GUSTAFSSON, A. BANE, P. MESCHAKS and J. E. RINGMAR

During their passage along the epididymal duct the spermatozoa undergo several changes (cf. Bishop, 1961) which may be related to changes in the composition of the epididymal plasma. It is known that in adult bulls the epididymal plasma changes characteristically in composition from one segment of the epididymal duct to another (Crabo & Gustafsson, 1964; Crabo, 1965).

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Employing the method previously developed by Crabo (1965), it was decided to perform analyses of the epididymal plasma of bull calves before or at the time when spermatozoa appear in the epididymal duct, and to compare the composition of the epididymal plasma in bull calves with that in adult bulls.

The material consisted of eighteen bull calves aged from 209 to 242 days, of Swedish Red and White breed, and crosses between this breed and

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R. S. Jeyendran, H. H. Van der Ven, M. Perez-Pelaez, B. G. Crabo and L. J. D. Zaneveld

Summary. The objective of this study was to develop a relatively simple test to evaluate the functional integrity of the membranes of human spermatozoa. As in some other species, human spermatozoa 'swell' under hypo-osmotic conditions due to the influx of water and the expansion of the membranes. A mixture of equal parts of fructose and sodium citrate (150 mosmol) with calculated ionic strength of 0·15 resulted in a maximal number of clearly identifiable swollen spermatozoa. Only small variations were seen when different aliquants of the same semen samples were separately evaluated. A high correlation (r = 0·94) was obtained between expected and observed values of swollen spermatozoa when known amounts of heat-treated spermatozoa, unable to undergo swelling, were added to untreated spermatozoa. A good correlation (r = 0·90) was also observed between the % spermatozoa in a semen sample that were capable of undergoing swelling and the % of denuded hamster oocytes that were penetrated by capacitated spermatozoa from the same semen sample. By contrast, the correlations between % sperm swelling in ejaculates and % normal sperm forms, % motile spermatozoa and % spermatozoa that do not stain with eosin-Y (supravital stain) in the same ejaculates were 0·30,0·61 and 0·52, respectively. Therefore, the hypo-osmotic swelling technique to evaluate the functional integrity of the sperm membrane appears to give high repeatability and accuracy and is closely correlated to the in-vitro fertilizing ability of spermatozoa. It may be a useful addition to the standard semen analysis.