Reproductive function of Hungarian Mangalica boars: effect of seasons

Seasonal changes in testis volume, testosterone (T) productivity (GnRH test) and semen characteristics of Mangalica boars were studied. The biggest testis volume was measured in autumn and the smallest in winter. Significant differences were demonstrated between autumn-winter (P = 0.012) and autumn-spring (P = 0.015) in testis volume. The highest basic T concentration (Tb) was observed in autumn and the lowest in summer. The provoked T concentration (Tincr) was significantly higher in autumn than in spring (P = 0.0007). A strong correlation was observed between T concentrations and testis volume in spring. The highest ejaculate volume was measured in winter while the lowest in autumn. Significant differences were found in semen concentration as well as in the total number of spermatozoa per ejaculate between seasons. The highest number of abnormal sperm cells was observed in spring while the lowest in summer. It can be concluded that the ejaculate of the Mangalica breed tends to be of lower volume and higher sperm concentration as compared to most pig breeds. Seasonal differences could be observed in testicular measurements, testosterone production capacity and sperm morphological features; however, sperm motility remained constantly high during the study.     

Differences in reproductive pattern between wild and domestic rams are not associated with inter-specific annual variations in plasma prolactin and melatonin concentrations

This study compares morphological changes, performed with real-time transrectal ultrasonography, of testis and accessory glands and seasonal endocrine changes in a wild (Mouflon) and a domesticated (Spanish Merino) breed of sheep. In Mouflons, the maximum plasma testosterone concentrations, testicular diameter and vesicular gland size occurred synchronously during autumn (P < 0.001). In Merino rams the highest circulating levels of testosterone (P < 0.05) and maximum testicular diameter (P < 0.001) occurred during summer, with no seasonal variations in vesicular and bulbourethral glands. The seasonal changes in plasma concentrations of prolactin were not correlated with annual variations in testicular and glandular size in neither wild nor domestic species. No differences were observed between both species for the seasonal pattern of prolactin secretion and mean amplitude of melatonin. Wide differences in reproductive patterns between wild and domestic types of rams do not appear to be attributed to seasonal changes in prolactin and melatonin secretion.     

Vascularization and Production of Mitogenic Factor(S) in Domestic Cat (Felis catus) Testes: Preliminary Results

Some male seasonal breeders are known to undergo testicular growth and regression throughout the year. The periodic and fast physiological development and regression processes, with simultaneous fast changes in blood flow, appears to be stimulated by angiogenic substances and inhibited by anti-angiogenic factors. The objective of this study was to evaluate the effect of season on microvascularization and production of angiogenic factors by testicular tissue, using the male cat as a model. Monthly, testes were collected post mortem from domestic stray cats, for histology and for tissue culture. Testis explants were cultured for 18h and conditioned media were tested for their ability to stimulate mitogenesis of bovine aortic endothelial cells (BAEC). Vascular endothelial growth factor (VEGF) was used as a positive control. Proliferative response of BAEC to samples was evaluated by determining the number of cells in each well using a Neubauer chamber. Vascular density was assessed by a computerized image analysis ('Scion Image, NIH, USA') based on the total histological area occupied by the vascular lumen. Percentage data subjected to arcsine transformation were analysed by one-way ANOVA and post-comparison tests. There was a significant increase in microvascular areas in November ( P  < 0.001), when compared to the remaining months (LSD test). All cat testicular tissue showed the capability to increase BAEC proliferation, when compared to negative controls, throughout the study. However, there was an increase in BAEC mitogenesis in February ( P  < 0.05) and November ( P  < 0.001), when compared to the other months. Our data show that cat testicular tissue from different periods of the year stimulates angiogenic factor(s). Furthermore, changes in angiogenesis may play a role on vascular growth and regression of the testes during the breeding and non-breeding season in the male cat.     

Effects of the GnRH antagonist acyline on the testis of the domestic cat (Felis catus)

The aim of this study was to describe the effects of a single dose of the gonadotrophin releasing hormone (GnRH) antagonist acyline on testicular characteristics of the domestic cat. Twelve mature cats were orchidectomised unilaterally (right testis) on Day -7 (n=7) or Day 15 (n=5). On Day 0, 330 μg/kg acyline was administered s.c. to all the animals. Left orchidectomy was carried out on Day 15 (n=2), Day 30 (n=4) and Day 60 (n=6). Sperm were recovered from the epididymis and the testes were evaluated grossly, histologically and immunohistochemically. Significant differences (P<0.05) were found between days for epididymal sperm motility, vigor, abnormal morphology, germinal epithelium height, spermatocytes, spermatids, spermatozoa, lumen and cellular debris. Conversely, no significant differences were found for gross testicular and tubular characteristics, spermatogonia, Sertoli and Leydig cells and intertubular compartments. It was concluded that a single dose of acyline reversibly impaired spermiogenesis, spermatocytogenesis and sperm motility for 2 weeks.     

Use of cyproheptadine to control urine spraying and masturbation in a cat

A 5-year-old male domestic longhair cat was examined because of urine spraying and masturbation. The cat had sprayed urine from the time it was acquired as a stray 4 years earlier. The cat was cryptorchid, and at 1 year of age, the scrotal testicle was removed. The cryptorchid testicle was surgically removed several months later; however, urine spraying and masturbation persisted. A diagnosis of territorial marking and separation anxiety was made. Serum testosterone concentration was within the reference range for sexually intact male cats. Treatment included behavior modification and administration of cyproheptadine (2 mg, p.o., q 12 h), which has been shown to have antiandrogenic effects in other species. Frequency of urine marking and masturbation decreased, along with serum testosterone concentration. The cat continued to do well as long as medication was given consistently. Eventually, the cat underwent a laparotomy for removal of remnant testicular tissues but was then lost to follow-up.     

Testosterone serum profile,semen characteristics and testicular biometry of Mangalarga Marchador stallions in a tropical environment

This study was conducted to characterize the daily profile of testosterone secretion and its mean concentrations in the four seasons as well as to evaluate the semen characteristics and testicular biometry of Mangalarga Marchador stallions throughout the year in a tropical region. Three stallions were submitted to semen collections and evaluation of testicular biometry every 14 days along a year. Blood samples were collected once at the middle of each season, in a 20‐min interval during 24 hr in order to evaluate the testosterone secretion profiles among seasons. Testosterone concentrations along the day were higher at the beginning of the afternoon (from 12:00 to 15:00 hr), but a circadian secretion was not clearly observed. Mean testosterone concentrations did not differ among seasons (p > .05), but a pattern of secretion along the day showed variations with higher concentrations in the afternoon during the winter. Ejaculate volume was higher during summer; however, sperm motility decreased in summer and spring. Total sperm in ejaculate, sperm morphology and testicular biometry kept constant along the year showing no differences among the seasons. The results demonstrated that in a tropical region, reproductive aspects of stallions did not show a clearly defined seasonal variation, and months of autumn and winter were not unsuitable for reproduction of the males.     

Androgen concentration in the blood and spermatogenic function of tom cats during the breeding season

In order to better understand androgen secretion in the tom cat during the breeding season, observations of diurnal changes in peripheral blood testosterone (T) levels, were made and the relation between androgen levels (androstenedione (A), 5 alpha-dihydrotestosterone (DHT). T) in testicular and peripheral vein blood was borne. Histologic examinations of the testis were also performed to investigate spermatogenic function. The 9 tom cats used in these studies were 2-3 years old, weighed 3.5-4.0 kg, and were raised in a room under natural lighting. As a result, diurnal peripheral blood T levels in the tom cat fluctuated greatly according to each individual, but since no definite trend could be identified, secretion was determined to be episodic. Although fairly large differences in the 3 types of androgen were observed in testicular vein blood depending on the individual animal, levels on the left and right were almost equal. Moreover, a correlation was found between the testicular and peripheral vein blood concentrations of the 3 androgens (A: p less than 0.01; DHT: p less than 0.05; T: p less than 0.01). Therefore, secretion of testicular androgen can be inferred by measuring peripheral blood androgen levels. Testicular histological examination revealed active spermatogenesis in all of the cats, and no significant inter individual differences were detected in either seminiferous tubule diameters or any of the various germ cells.     

The Annual Testicular Cycle of the Domestic Quail (Coturnix coturnix japonica)

A morphometric study was conducted on the testis of the domestic quail Coturnix coturnix japonica to determine testicular kinetics. We investigated the variability along the year of testicular parameters such as seminiferous tubule diameter, germinal epithelium height and amount of meiotic figures of maturing spermatids in the seminiferous epithelium and of sperm in the tubular lumen. The results of morphometric analysis showed the occurrence of an annual testicular cycle defined by four distinct phases: a resting phase (at the end of summer), a recrudescence phase (in the fall), a proliferative phase (at the end of winter and beginning of spring), and a regression phase (spring and summer). We also observed that the testes of adult quails present elevated and maximal spermatogenic activity in fall-winter (short-day period) and at the beginning of spring, respectively, and lower values in spring and summer (long-day periods), with minimum values at the end of summer.     

The Effect of Season on Semen Characteristics and Freezability in Bos indicus and Bos taurus Bulls in the Southeastern Region of Brazil

The objectives of this study were to evaluate the effects of season in southeast of Brazil comparing genotypes on semen characteristics, freezability and peripheral plasma concentrations of testosterone. Ejaculates of five Bos indicus bulls and six Bos taurus bulls were evaluated over a period of 27 months, which was divided into winter (July, August, September), spring (October, November, December), summer (January, February, March) and autumn (April, May, June). Semen was evaluated according to standard procedures for ejaculate volume, sperm concentration, gross-motility, progressive motility and sperm morphology. After preparing and freezing the ejaculates according to commercial procedures, the straws were stored in liquid N₂ until post-thaw evaluation. Ejaculate volume, sperm concentration, gross-motility, progressive sperm motility, vigor and morphological sperm defects were significantly influenced by season and genotype (p < 0.05). Heat tolerance was better in B. indicus bulls than in B. taurus bulls characterized by lower values of sperm abnormalities throughout the observation period. The highest values were recorded for abnormal heads followed by cytoplasmatic droplets in B. taurus bulls. The proportion of ejaculates which were eliminated before freezing for reasons of bad quality was lower in the B. indicus bulls. Temporal changes in peripheral plasma testosterone concentrations were higher in B. indicus bulls than in B. taurus bulls not revealing seasonal influences. The results of this study show clear genotype differences regarding semen quality. Freezability of B. taurus semen varies considerably throughout the year, leading to a high proportion of eliminated ejaculates. Collecting semen from B. taurus bulls during the summer in an artificial insemination centre may not be profitable.     

Changes in the Immunolocalization of Steroidogenic Enzymes and the Androgen Receptor in Raccoon (Procyon lotor) Testes in Association with the Seasons and Spermatogenesis

The raccoon is a seasonal breeder with a mating season in the winter. In a previous study, adult male raccoons exhibited active spermatogenesis with high plasma testosterone concentrations, in the winter mating season. Maintenance of spermatogenesis generally requires high testosterone, which is produced by steroidogenic enzymes. However, even in the summer non-mating season, some males produce spermatozoa actively despite low plasma testosterone concentrations. To identify the factors that regulate testosterone production and contribute to differences in spermatogenetic activity in the summer non-mating season, morphological, histological and endocrinological changes in the testes of wild male raccoons should be known. In this study, to assess changes in the biosynthesis, metabolism and reactivity of testosterone, the localization and immunohistochemical staining intensity of four steroidogenic enzymes (P450scc, P450c17, 3βHSD, P450arom) and the androgen receptor (AR) were investigated using immunohistochemical methods. P450scc and P450c17 were detected in testicular tissue throughout the year. Seasonal changes in testosterone concentration were correlated with 3βHSD expression, suggesting that 3βHSD may be important in regulating the seasonality of testosterone production in raccoon testes. Immunostaining of P450arom and AR was detected in testicular tissues that exhibited active spermatogenesis in the summer, while staining was scarce in aspermatogenic testes. This suggests that spermatogenesis in the raccoon testis might be maintained by some mechanism that regulates P450arom expression in synthesizing estradiol and AR expression in controlling reactivity to testosterone.     

Plasma Hormone Levels and Semen Quality in Male Cats during Non-Breeding and Breeding Seasons

Female cats are known to be seasonal breeders and male cats annual breeders. Despite this, there are limited data on the influence of breeding season (BS) on hormone concentration and semen quality in the male cat. This study compared plasma concentrations of LH and testosterone (T), and semen quality during the non-breeding season (NBS) and BS in five male cats subject to natural hours of daylight but a constant environmental temperature. Plasma LH and T concentrations were higher during the BS in 2/35 and 3/5 cats, respectively, although when comparing both hormones combined, values were higher during the BS than the NBS in all cats (p < 0.01). There were no significant differences in the percentage of abnormal sperm between the cats. Overall, semen quality was superior during the BS with larger semen volume in 2/5, sperm motility in 2/5 and sperm viability in 3/5 cats. Although there was a clear seasonal effect on hormone secretion and semen quality, during the NBS all cats were likely to have been fertile.     

Cryopreservation of cat testicular tissues: effects of storage temperature, freezing protocols and cryoprotective agents

Cryopreservation of testicular tissue has become a part of gamete preservation in wild animal post-mortem. Using domestic cats as a model for wild felids, this study aimed to (i) investigate the effect of temperature for testicular tissue storage on sperm quality; (ii) compare efficiency of freezing protocols; and (iii) evaluate properties of cryoprotective agents to protect testicular sperm quality. A pair of testes from each cat (n = 9) was cut into four pieces. Three randomly selected pieces were allocated to be (i) fresh controls; (ii) stored at 4 °C for 24 h; and (iii) stored at room temperature (28 °C) for 24 h. After storage, the testicular tissue from each group was cut into 10 small pieces. One piece was assigned to be a control while the others were assigned to three freezing protocols; -80 °C (n = 3), vitrification (n = 3) or two-step freezing (kept above liquid nitrogen vapour for 10 min and submerged in liquid nitrogen) (n = 3). Each of three pieces was frozen using dimethyl sulphoxide (DMSO), ethylene glycol (EG) or DMSO combined with EG. Sperm membrane (SYBR-14/EthD-1) and DNA (acridine orange) integrity were evaluated before and after cryopreservation. The storage of testicular tissue at room temperature decreased the percentage of sperm with intact membrane in fresh tissue (59.5 ± 30.5 vs 87.9 ± 7.0%, p < 0.05). DNA integrity was decreased after 24-h storage either at 4 °C or room temperature (p < 0.05). The two-step freezing resulted in a higher percentage of sperm with intact plasma membrane than the other techniques. Dimethyl sulphoxide, EG and DMSO combined with EG provided similar protection for the sperm membrane and DNA from cryodamages. In conclusion, storage of testicular tissue at 4 °C is necessary to maintain sperm membrane integrity during transportation of tissue for cryopreservation in the freezing laboratory. The results provide information for male gamete rescue in felid particularly when they die unexpectedly in the field where freezing facilities are not well equipped.     

The relationship of plasma leptin concentration and puberty in Holstein bull calves (Bos taurus)

The objective of this experiment was to study the changes of plasma leptin concentration during puberty and its relationship with testosterone level and testis dimensions in Holstein bull calves. Six Iranian Holstein bull calves with approximately 6 months of age were used. Semen evaluation was conducted at 1‐month interval to determine the puberty state. To detect the plasma leptin and testosterone changes, blood samples were collected from the jugular vein during pre‐puberty (6–7 months of age), puberty (8–9 months of age) and post‐puberty (10–11 months of age). In addition, body weight (BW), body condition score (BCS) and testicular width and length were measured at 3‐week intervals. The effects of time (age) on total sperm number and percentage of progressive motility of sperm, plasma concentration of leptin and testosterone, amplitude and frequencies of testosterone, BW, BCS, testicular dimensions were significant. Sperm number and progressive motility during post‐puberty were higher than those during puberty and pre‐puberty. Plasma concentration of leptin during the pre‐puberty was higher than those during puberty and post‐puberty (p < 0.01). Mean plasma testosterone concentrations during puberty were higher than those during pre‐puberty (p < 0.05). BW, BCS and testicular dimensions consistently increased throughout the trial. Results indicated that in growing bull calves, plasma concentrations of leptin decreased during puberty, while circulating testosterone increased.     

Seasonal Changes in Testicular Cell Morphology in Domestic Male Cats (Felis catus)

The aim of this study was to asses the variation in the morphology of the seminal epithelium in relation to natural photoperiod in male cats. Tom cats (n = 240) were castrated every other week throughout the year. Each testis was fixed in Bouin's solution and cut into sections. The percentage of tubules with round spermatids (RS), elongated spermatids (ES), tailed spermatids (TS), mature spermatids (MS) and the number of Sertoli cells (SC) and Leydig cells (LC) were recorded in each sample. Testicles from males during short days (SHD) had a higher percentage of tubules with RS and ES compared to testicles from males during long days (LHD, 31.3 ± 0.6 vs 2.1 ± 0.6%, p < 0.001; 30.9 ± 0.7 vs 11.0 ± 0.7%, p < 0.001). Conversely, testicles from males during SHD had a lower percentage of tubules with TS and MS compared to testicles from males during LHD (24.5 ± 0.8 vs 29.7 ± 0.8%, p < 0.01; 13.1 ± 1.2 vs 57.0 ± 1.2%, p < 0.01). Furthermore, testicles from males during SHD had a higher number of SC and lower number of LC compared to testicles from males during LHD (11.4 ± 0.1 vs 8.0 ± 0.1%, p < 0.01; 19.2 ± 1.0 vs 38.0 ± 1.0%, p < 0.01). In conclusion, there are seasonal changes in testis cell morphology in the tom which may be related to seasonal sperm production.     

Neuropathologic findings of bromethalin toxicosis in the cat.

Ten random source male domestic shorthair cats, 2 to 6 years old and 3.0-4.4 kg body weight, were each given a single oral dose (1.5 mg/kg) of bromethalin (cat Nos. 1-5) or bait vehicle carrier (cat Nos. 6-10). Bromethalin-dosed cats developed a toxic syndrome characterized by ataxia, focal motor seizures, vocalization, decerebrate posture, decreased conscious proprioception, recumbency, depression, and semicoma. Bromethalin-dosed cats were euthanatized if seizure activity or hindlimb paralysis developed. Survival times were 48 hours (cat No. 1), 89 hours (cat No. 2), 90 hours (cat No. 3), and 97 hours (cat No. 4). Control cats (cat Nos. 6-10) and one bromethalin-dosed cat (cat No. 5) were euthanatized on day 20 after dosing. Spongy change (edema--characterized by the formation of vacuoles in extracellular spaces and myelin lamellae), hypertrophied fibrous astrocytes, and hypertrophied oligodendrocytes were observed in the white matter of the cerebrum, cerebellum, brain stem, spinal cord, and optic nerve of all bromethalin-dosed cats. Spongy change occasionally extended into contiguous cerebellar Purkinje cell layer and cerebral cortical gray matter. The severity of lesions varied among cats but was most pronounced in cat No. 5 (480 hours after dosing). A leukocytic inflammatory response, gitter cell (macrophage) response, or axonal degeneration was not observed in the vacuolated areas. Ultrastructural findings included separation of myelin lamellae at the interperiod lines with the formation of intramyelinic vacuoles (intramyelinic edema), rupture and coalescence of intramyelinic vacuoles into larger extracellular spaces (spongy change), and pronounced cytosolic edema of astrocytes and oligodendroglial cells.     

Induction of sexual activity in lactating anovulatory female goats using male goats treated only with artificially long days

Two experiments were conducted to determine the response of Creole male goats treated with long days and melatonin implants, and the response of the anovulatory does to male effect using males treated only with artificially long days. All animals were allocated to open sheds. In Exp. 1, one group of males was under natural photoperiod (CG; n = 7); the second group was submitted to 2.5 mo of long days followed by the insertion of two s.c. melatonin implants (LD+MEL; n = 7); the third group was subjected only to 2.5 mo of long days (LD; n = 7). Testicular weight was measured every 2 wk. Plasma testosterone concentrations were determined weekly. A treatment x time interaction was detected (P < 0.001) for testicular weight and plasma testosterone concentration. In the LD+MEL and LD groups, testicular size and plasma testosterone levels varied in a similar way, but differed from those observed in CG (P < 0.001). In this latter group, testicular weight displayed seasonal variations and peaked in June, whereas in treated groups this peak occurred in March. In CG, testosterone varied in a seasonal manner and plasma concentrations increased in June and remained elevated throughout the study. In experimental groups, testosterone increased in February and peaked in March. In Exp. 2, one group of males was left under natural photoperiod (CG, n = 5) and the other one was submitted to 2.5 mo of artificially long days (LD, n = 4). On March 16, two control and two treated males were put in contact with 20 and 19 females, respectively. Sexual behavior of the bucks was observed during the 5 d following male introduction. Progesterone assays and estrous behavior were used to determine ovarian and behavioral responses of the females to teasing. The anogenital sniffing, nudging, and mount instances registered in LD-treated males were greater than those observed in CG (P < 0.05). Of the does exposed to CG, none ovulated and only two of 20 females displayed estrous behavior. All does in contact with LD-treated males ovulated and showed at least one estrous behavior during the 15 d following joining (P < 0.001). These results indicate that the sexual activity of male goats from subtropical latitudes can be induced using only artificially long days. In addition, males treated in this way are capable of stimulating sexual activity in anovulatory females by the male effect.     

Morphofunctional evaluation of the testicle and the spermatogenic process of adult white-eyed parakeets (Aratinga leucophthalma MULLER, 1776) during the different seasons of the year

In this experiment, testicle fragments of 14 adult White-eyed Parakeets (Aratinga leucophthalma) were evaluated as for their seasonal reproductive activities using the following quantitative parameters: average thickness of the testicular tunica albuginea, volumetric proportion of tubular and extratubular compartments, average diameter of the seminiferous tubules and corporal weight. Parameters were created for qualitative evaluations of the degree of spermatogenic development. In this experiment, all the animals were distributed into four groups, and their testicular fragments were collected during the middle of summer, fall, winter and spring. The animals were submitted to volatile general anaesthesia, and a biopsy was made by celioscopy. The fragments collected were processed histologically. The slides were prepared and later evaluated by using an optical microscope. The average seasonal values of the corporal weight increased, starting in the winter and reaching the peak during the spring. A seasonal testicle cycle was observed, because, in the spring, the testicles showed values for the quantitative and qualitative parameters of spermatic production compatible with the period of greater activity, while the opposite thing happened during the fall. Our data indicate that the parameters of sperm production may be correlated with daily light rather than with air humidity.     

Effect of divergent selection for testosterone production on testicular morphology and daily sperm production in boars

The objective of this study was to characterize correlated responses in testicular morphology and daily sperm production to divergent selection for testosterone production. Duroc boars from high and low lines (HTL and LTL, respectively) divergently selected over 10 generations for testosterone production in response to a GnRH challenge followed by random selection were used. Testicular tissues were sampled from all available males of generation 20 (HTL, n = 46; and LTL, n = 13). Volume densities for Leydig cells, seminiferous tubules, and Sertoli cells were estimated along with sperm production. The HTL boars had greater volume densities of Leydig cells than did LTL (P < 0.01). Volume density of seminiferous tubules tended to differ between lines (P < 0.07), but Sertoli cell volume densities did not differ (P < 0.27). Sperm production traits, adjusted for age, did not differ significantly between lines. Body, testicular, and epididymal weights were recorded for boars from HTL (n = 82) and LTL (n = 44) from generations 20 and 21. After adjustment for BW, average paired testicular weights for HTL and LTL were 417 and 457 g (P < 0.01), respectively. Epididymal weights, adjusted for BW, were heavier for HTL (P < 0.01) than for LTL. To demonstrate that the selection lines still differed for testosterone production, lines were evaluated in generation 21. Endogenous testosterone production of the HTL (n = 54) and LTL (n = 44) testosterone production line averaged 49.0 ng/mL and 27.8 ng/mL (P < 0.01), respectively. Plasma FSH concentrations did not differ between lines (P < 0.30). Selection for testosterone production in response to a GnRH challenge was an effective method of changing testosterone concentrations, testicular size, epididymal weight, and volume density of Leydig cells. However, daily sperm production per gram of testes was unchanged. Based on the results of this study, selection for testosterone production is not recommended as a method of increasing sperm production in pigs.     

Effects of Intratesticular Injection of 70% Glycerin on Stallions

The removal of endogenous germ cells of recipient stallions is a key step to produce donor germ cell-derived sperm using the germ cell transplantation technique. Six Thoroughbred stallions were divided into a treatment (n = 3) and a control group (n = 3), and 70% glycerin (1, 2, 3-trihydroxypropane, 40 mL per testis) or phosphate-buffered saline, respectively, was locally injected into testes. General semen evaluation, libido, and testicular volume were performed weekly from 3 weeks before to 10 weeks after treatment. The number of round germ cells in the ejaculate was counted using a hemocytometer. The hematoxylin and eosin staining was performed on the cross sections of testicular tissue obtained 11th week of treatment. Plasma testosterone levels in blood collected weekly were measured using a colorimetric competitive enzyme immunoassay kit. The sperm number was significantly lower than that of the control group at 5 and 10 weeks after glycerin injection. No differences in the status of spermatogenesis in the cross sections of seminiferous tubules and testicular volume were found between the two groups. The 70% glycerin-treated stallions had reduced total and progressively motile sperm and exhibited a significantly higher population of round germ cells in the ejaculate. Testosterone levels, testicular volumes, and libido of stallions were not significantly different between the groups. In conclusion, although intratesticular injection of 70% glycerin may have caused disassociation of some germ cells in the seminiferous tubules for several weeks, it did not significantly ablate germ cells in the tubules at 11 week in stallions.     

Evaluation of a Burdizzo Castrator for Neutering of Dogs

A Burdizzo castrator was evaluated for the neutering of dogs. Histological and morphological changes of spermatic cells and peripheral serum testosterone after challenge with a GnRH-analogue (gonadorelin) were assessed. There was a control group (G1), a surgically castrated group (G2) and a Burdizzo group (G3) divided in two, G3a receiving two crunches in each spermatic cord and G3b receiving one crunch in each spermatic cord. Sixteen days after application of the Burdizzo blood samples were taken from the dogs at 30 min interval during 2 h; after the second sample the dogs were treated with 1 mug/kg body weight of gonadorelin i.v. The same protocol of gonadorelin challenge was performed in G1 and G2 dogs. The G2 dogs were surgically castrated after the second blood sample, before the gonadorelin treatment, and the G1 dogs after the last blood sample. The excised gonads were examined histologically, and sperm smears were prepared from the caudae epididymidis. The testes and plexus pampiniformis of the G1 and G2 dogs had a normal histological appearance, and they had morphologically normal epididymal sperm cells. In all G3 dogs, there was an acute fibrosis with an inflammatory reaction in the plexus pampiniformis. The testes from the G3a dogs showed diffuse areas of infarction and degeneration of the parenchyma. Similar but less diffuse lesions were seen in group 3b dogs. The deferent ducts from all G3 dogs showed vasitis and/or sperm granulomas. Azoospermia or sperm malformations were observed in the epididymal smears from the G3 dogs. Testosterone concentration in the G1 dogs increased after gonadorelin application (p < 0.0001). The G2 dogs had basal testosterone levels after castration (p < 0.001) and did not respond to gonadorelin. Groups 3a and b showed a slight but non-significant increase in testosterone concentration after gonadorelin challenge, supposedly due to the reduction of testicular blood flow and loss of testicular interstitial tissue.