Immunological castration temporarily reduces testis size and function without long-term effects on libido and sperm quality in boars

The objective was to determine the effects of immunization against gonadotropin-releasing hormone on reproductive characteristics in boars. A total of 72 boars were used in a randomized design with three treatments: single immunization (SI) (10 weeks of age) or double immunization (DI) (10 and 15 weeks of age) with Improvest® and intact controls (no Improvest®; CNT) (n=24/group). At 10, 15, 20, 25 and 40 weeks of age, blood was collected and serum harvested to evaluate testosterone concentrations. Testosterone concentrations were less for DI boars compared with CNT boars and SI boars at 20 and 25 weeks (P<0.001), but not at 40 weeks of age. At week 25, 18 pigs (n=6/group) were sacrificed and testes were removed, weighed and measured, and seminiferous tubules were examined and scored using histological slides of testes parenchyma. A sample of neck fat was assessed for boar taint aroma. All testicular measurements and weights and seminiferous tubule scores were less for DI boars compared with SI and CNT boars (P<0.001). More (P<0.05) SI and CNT boars had detectable boar taint aroma than DI boars. Libido was assessed at 32, 36, 47, 60 and 63 weeks of age and semen collected at 60 weeks of age was analyzed for indicators of quality. There were no effects of treatment (P=0.41) or treatment by week (P=0.71) on libido. Semen volume, gel weight and total number of sperm cells, determined in a subset of boars (n=3/treatment), were not different among treatments. Sperm concentration was greater for DI than SI (P=0.01), and tended to be greater for DI compared with CNT (P=0.10). Sperm motility tended to be greater for DI boars compared with CNT boars (P=0.066). In conclusion, our results show that there are no long-term effects of immunocastration on reproductive characteristics in boars.     

Pituitary gonadotropins, hypothalamic gonadotropin-releasing hormone, and testicular traits of boars exposed to natural or supplemental lighting during pubertal development

Seventy crossbred boars were reared under natural (30 lux) or supplemental lighting (1000 lux) beginning at 4 wk of age. Boars received supplemental lighting from six 40-watt fluorescent bulbs between 0530 and 2030 h. Five boars from each treatment were killed at 67, 91, 119, 155, 182, 210, or 246 days of age. No differences (p greater than 0.05) in pituitary concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and prolactin (PRL) were found between treatment groups at any age. Total pituitary content of LH, FSH and PRL increased as boars became older, but when expressed as hormone concentration, only PRL increased with age. Content of gonadotropin-releasing hormone (GnRH) in the pituitary stalk-median eminence, preoptic area, and hypothalamus proper was similar (p greater than 0.05) between treatments. When GnRH contents were totaled and combined for the treatment groups, it was found that GnRH content increased (p less than 0.05) as boars became older. No differences (p greater than 0.05) were observed in testicular volume percentage of seminiferous tubules and tubular diameter between lighting treatments. These data demonstrate that the supplemental lighting does not influence puberty in boars by altering hypothalamic content of GnRH or pituitary stores of LH, FSH, and PRL.     

In vitro fertilization of porcine oocytes with fresh and frozen-thawed ejaculated or frozen-thawed epididymal semen obtained from identical boars

The objective of this study was to compare the in vitro fertilizing capacity of porcine spermatozoa from fresh and frozen-thawed semen and frozen-thawed epididymal spermatozoa obtained from identical boars. Prior to IVF, fresh spermatozoa were capacitated in TCM 199. Frozen semen samples were stored in 0.25-ml plastic straws using a lactose/glycerol/orvus-es-paste extender. Cumulus-oocyte-complexes (COC) obtained from superovulated prepuberal gilts were fertilized in vitro within 2 h after aspiration with one of the semen samples. After final dilution for IVF, frozen-thawed epididymal semen samples showed motility rates (72.2 +/- 5.6%) similar to those of spermatozoa in fresh semen (76.4 +/- 4.5%), while sperm motility decreased in frozen-thawed ejaculated semen (40.2 +/- 9.4%). Considerable individual differences in sperm motility between boars were observed for ejaculated semen but not for epididymal semen. Enhanced fertilizing capacity of frozen-thawed epididymal spermatozoa was confirmed by pronucleus formation and cleavage rates, with significantly more embryos developing to the 2- and 4-cell stages compared with the groups fertilized with fresh or with frozen-thawed ejaculated semen (59.7 vs 14.6 and 16%). In conclusion, consistent in vitro fertilization rates with minimal semen variability are obtained using frozen-thawed epididymal semen. Following a modified freezing protocol, epididymal spermatozoa can easily be frozen in small containers for IVF, with higher resultant motility and fertilization rates than with ejaculated semen.     

Porcine sperm viability, oocyte fertilization and embryo development after staining spermatozoa with SYBR-14

The objective of these experiments was to determine the efficacy of the new membrane permeant nucleic acid stain, SYBR-14, for assessing boar sperm viability and to determine it's effect on fertilization and early embryonic development using the pig as a model. We examined the staining patterns of SYBR-14 and another vital stain, Hoechst 33342, both in combination with the dead cell stain, propidium iodide (PI), to quantify the proportion of living and dead spermatozoa in ejaculated and epididymal semen. Flow cytometry analyses of semen from 4 boars revealed significant differences among boars for the proportion of SYBR-14-stained spermatozoa in both epididymal and ejaculated samples, but not for Hoechst 33342 or PI stained spermatozoa. Gilts were inseminated with unstained spermatozoa or spermatozoa stained with 2 levels of SYBR-14 or 2 levels of the reference stain, Hoechst 33342. Embryos recovered at 42 to 48 h postinsemination were morphologically evaluated, and only 4 to 8-cell embryos were continued in culture. Overall, fluorescent staining of boar spermatozoa with SYBR-14 or Hoechst 33342 neither affected their ability to fertilize oocytes, nor the developmental competence of the resultant embryos.     

Return of testicular function after vaccination of boars against GnRH: consequences on testes histology

Traditionally, male pigs are surgically castrated without anaesthesia to avoid later occurrence of the sex odour of androstenone in the carcass. Active immunization against gonadotropin-releasing hormone (GnRH) is a painless alternative which inhibits LH and thus steroidogenesis in the Leydig cells. In a preceding study we clarified the return of Leydig cell function after the last dose of antigen by measuring hormones, and found a considerable variation (10 to 24 weeks) till return of their function (testosterone 0.5 ng/ml blood plasma). The present paper analyses histological data on testes characteristics of the same six boars at an age of 52 weeks (26 weeks after last immunization). Data were compared to another four boars which were not immunized but slaughtered at the same age. Testis weight was related to the concentration of testosterone in blood. In boars, that first returned to testicular function, testis weight even exceeded those in controls probably due to rebound phenomena. Differences in testis weight were mainly due to differences of Leydig cell content of cytoplasm, and less to the size of nuclei. Additionally, the height of seminiferous epithelium was slightly dependent on testosterone concentrations and contributed moderately to differences in testis weight. Altogether, normalization of testicular function, even after return to steroidogenesis, requires another 13 weeks.     

The effect of lutalyse on the training of sexually inexperienced boars for semen collection

The objective was to determine if i.m. treatments of lutalyse (PGF2alpha; dinoprost tromethamine salt) expedited the training of sexually inexperienced boars for semen collection. Lean-type, terminal-line boars (n = 40; 177.4 +/- 2.4 day of age and 112.8 +/- 2.0 kg body weight) that had not previously experienced natural mating were utilized. Boars were moved individually twice weekly for 6 weeks (total of 12 training sessions) to a semen collection room equipped with an artificial sow. Upon entering the semen collection room, boars received i.m. treatments of either deionized water (4 ml, n = 10) or lutalyse at doses of 5 mg (n = 10), 10 mg (n = 10), or 20 mg (n = 10), and subsequently received a libido score of 1-5 (1 = no interest in the artificial sow; 5 = mounting artificial sow and allowing semen collection). The percentages of boars successfully trained for semen collection during the experimental period were similar (P > 0.05) for controls (20%) and boars receiving 5 mg (30%), 10 mg (20%), or 20 mg (10%) of lutalyse. Average libido score for boars receiving 10 mg lutalyse (2.35 +/- 0.08) was greater (P < 0.05) than for controls (2.14 +/- 0.06). In summary, lutalyse increased libido scores, but did not affect the number of boars trained for semen collection.     

Dietary l-arginine supplementation improves semen quality and libido of boars under high ambient temperature

l-Arginine is a nutritionally essential amino acid for spermatogenesis and plays versatile roles in animal health and can be utilized as a potential agent to improve reproductive performance of boars under high ambient temperature. The present study aimed to determine whether dietary l-arginine could alleviate heat stress-induced infertility in boars. In all, 20 boars (PIC 1040; 248.59±3.84 kg BW and 407.65±6.40 days of age) were selected and randomly assigned to four groups (group 0.0%, basal diet; group 0.6%, 0.8% or 1.0%, basal diet added with 0.6%, 0.8% or 1.0% l-arginine (wt:wt), respectively.) The four diets were made isonitrogenous by addition of appropriate amounts of l-alanine. Boars were pre-fed the corresponding experimental diet for 42 days. Then, the semen characteristics and libido were accessed for 6 weeks during the hot summer period (25.5° to 33.0°C). Results show that dietary l-arginine remarkably improved sperm motility, normality, total sperm number and effective total sperm number. Also, dietary l-arginine improved semen antioxidant capacity, such as decrease of malondialdehyde and 8-Hydroxy-2'-deoxyguanosine content in sperm (P<0.05), increase of the ratio of glutathione and oxidized glutathione, total antioxidant capacity, glutathione peroxidase and catalase activities in seminal plasma (P<0.05). Most of mitochondria contained intact ultrastructure in l-arginine-supplemented group which also accompany with higher ATP content than the 0.0% group. The boars fed 0.8% l-arginine show increased levels of estradiol-17β and testosterone and exhibit improved libido performance than boars in the 0.0% group. Adding dietary l-arginine linearly increased (P=0.002) nitric oxide content (as l-arginine increased). The scrotal surface temperature in the 0.6%, 0.8% and 1.0% group were decreased by 0.9°C, 0.9°C and 0.4°C, respectively, compared with the 0.0% group. l-Arginine levels caused linear effect on semen quality and antioxidant capacity, also caused quadratic effect on libido performance. During the hot summer months, the predicted optimal l-arginine levels for best semen quality and antioxidant capacity was 0.8% to 1.0% and for best libido performance was 0.8%. It can be concluded that l-arginine can be used as an effective agent to alleviate heat stress-induced infertility of boar, and that 0.8% to 1.0% can be considered as the optimum dosage.     

A Comparative Study on the Chemical Composition of Plasma from the Cauda Epididymidis,Semen Fractions,and Whole Semen in Boars

A comparative study was carried out on the chemical composition of plasma from the cauda epididymidis, semen fractions, and whole semen of boars. A total of 22 boars were used in this study. The boars, which ranged in age from 8 to 14 months, were of Swedish Landrace and Swedish Yorkshire breed. All boars used presented a normal semen picture. A dummy sow and an artificial vagina were employed for semen collection. The semen was collected as whole semen and as semen fractions in 10 nil volumes. The contents of the cauda epididymidis was removed post mortem. The following parameters were investigated: sperm concentration, dry weight of spermatozoa and of seminal plasma, osmotic pressure, sodium, potassium, chloride, inorganic phosphorus, calcium, magnesium, total protein, GOT, GPT, and alkaline phosphatase in seminal plasma. Paper electrophoresis was carried out on seminal plasma. Tlxe results of the analysis are summarized in Tables 1–6. The sperm concentration was approximately 3.2 mill./mm3 in the cauda epididymidis, 1 mill./mm3 in the sperm-richest fraction (II) and 0.25 mill./mm3 in whole semen. The dry weight (expressed in per cent dry matter) of spermatozoa was highest in the cauda epididymidis (25.47 %), showing a tendency to decreasing in semen fractions I—IV and was lowest in whole semen (15.29 %). The per cent dry weight in plasma was higher in the cauda epididymidis (4.56 %) than in semen fraction I (2.20 %). In semen fractions I—IV the per cent dry weight rose from 2.20 (U to 4.51 % and reached the level of approximately 3.80 % in the sperm-free fractions V—VII. The osmotic pressure was significantly higher in the cauda epidi-dymal plasma than in the whole seminal plasma or the seminal plasma fractions. The same phenomenon was observed in a boar where the cauda epididymal content was collected in vivo from a patent established fistula. There appears to be a connection between the per cent dry weight of spermatozoa and the osmotic pressure, which means that the per cent dry weight of the cauda epididymal spermatozoa decreases when mixed with the accessory gland secretions, which have a lower osmotic pressure. The fall in per cent dry weights is thought to be caused by an intake of water. The amount of sodium, chloride and magnesium was higher in ejaculated seminal plasma than in cauda epididymal plasma. The reverse was true for inorganic phosphorus and potassium. Moreover the sperm-free fractions contained more sodium, chlorides and magnesium than the sperm-containing fractions, while the concentration of potassium and inorganic phosphorus was comparatively higher in the sperm-containing fractions. A connection is apparent between sperm concentration and the potassium, inorganic phosphorus and magnesium levels. Statistical analysis of the values of chloride and magnesium revealed significant differences between individual boars for most of the semen fractions. The concentration of plasma proteins in the cauda epididymidis was approximately the same as in whole semen and in the semen fractions except for fraction I, which contained a relatively low concentration. As regards total protein there were significant differences between individual boars in most of the semen fractions as well. The paper electrophoretic pattern of epididymal plasma was different from that of semen plasma. Thus there were three or four distinct components in the cauda epididymidis numbered 1, 2, 3, and 4, and three distinct components in whole seminal plasma numbered 3, 4, and 5, while the sperm-richest semen fractions contained four components (2, 3, 4, and 5) and the others three components, namely 3, 4, and 5. The level of GOT was high in the cautlu cpiflidymill contents (99.1 i. u./ml) compared with that for whole seminal plasma (99.1 i.u/ml). In semen fractions there was a clear positive correlation between the level of GOT and the sperm concentration. The GPT concentration wis as a whole low and. in contrast to GOT. somewhat higher in the sperm-free fractions than in the sperm-containing fractions. The concentration of alkaline phosphatase was very high in cauda epididymal plasma (31,463 i. u./ml) as well as in the sperm-rich fractions (e.g. 7,096 i. u./ml in fraction II). Preliminary investigation has moreover revealed a very low alkaline phosphatase concentration in seminal plasma of vasectomized boars, which condition suggests thai the main origin for alkaline phosphatase in boars is the testis and epididymis.     

Semen quality of postpubertal boars during increasing and decreasing natural photoperiods

The present study analyses the effects of increasing and decreasing photoperiods on the semen quality of 20 selected postpubertal Landrace boars. The boars were exposed, throughout 75 days, to increasing and decreasing photoperiods of natural light, a constant temperature of 21 +/- 1 degrees C and 60-70% of humidity, fed with a nutritious diet and, submitted to a rhythm of semen collection of twice a week. During the last 2 weeks of each treatment, semen samples were analysed and the parameters measured were: ejaculate volume and pH, sperm concentration, sperm production and the number of semen doses per ejaculate, sperm vitality, sperm motility, osmotic resistance of spermatozoa and sperm morphology. The comparative analysis between increasing and decreasing photoperiods indicated that the semen quality of boars exposed to a decreasing photoperiod was reduced as a consequence of decreases in sperm concentration, sperm production and the number of semen doses. There was no difference between increasing and decreasing photoperiods in terms of sperm vitality and sperm motility, nor in the osmotic resistance of spermatozoa to isotonic and hypotonic media. The analysis of sperm morphology showed significantly lower frequencies of mature and immature spermatozoa with a distal cytoplasmic droplet, and significantly higher frequencies of immature spermatozoa with a proximal droplet in boars exposed to the decreasing photoperiod. These results indicate that the sperm quality of the selected boars decreased during decreasing photoperiods, in comparison with increasing photoperiods, mainly due to impaired testicular function.     

Pituitary and Leydig cell function in boars actively immunized against gonadotrophin-releasing hormone

Crossbred boars were (a) immunized against GnRH conjugated to human serum globulin (200 micrograms GnRH-hSG) in Freund's adjuvant at 12 weeks of age and boosted at weeks 18 and 20 (N = 10), (b) served as controls and received hSG only in adjuvant (N = 10), or castrated at weaning (N = 10). At 24 weeks of age (immediately before slaughter), the boars were challenged with saline or pig LH (1 microgram/10 kg body weight). After slaughter, fresh testicular fragments were incubated with pig LH (0.05 and 0.2 ng/2 ml medium) to assess the effects of immunization on Leydig cell function. Pituitary contents of LH and FSH, and testicular LH receptor content were also measured. The results indicated that plasma LH and testosterone concentrations, pituitary LH content, testicular LH receptor content, testis and sex accessory organ weights were significantly reduced in GnRH-immunized boars compared to hSG-adjuvant controls. However, plasma and pituitary FSH content were not affected by high antibody titres generated against GnRH. The testicular testosterone response to exogenous LH in vivo and in vitro was significantly reduced (P less than 0.05) in GnRH-immunized boars. These results indicate that active immunization against GnRH impairs pituitary and Leydig cell functions in boars.     

Peripheral plasma testosterone concentration and sexual behavior in young prenatally stressed boars

The purpose of this study was to examine the effects of stress induced physiological changes in the gestating sow on postnatal sexual and endocrine development of male offspring. Ten boars, ranging from 160 to 185 days of age, were randomly chosen from sows which had been maintained under either stress or control conditions during mid-gestation. Blood samples were collected weekly from each boar (minimum of four weeks) at 30 min intervals over a common six-hour period via an indwelling anterior vena cava cannula. Plasma testosterone concentrations were determined by radioimmunoassay. In order to ascertain degree of sexual behavior, boars were exposed weekly to gilts in estrus and a subjective score assigned. No differences (P>.10) were found between prenatally stressed and control boars in overall mean testosterone concentration or libido score. A significant (P<.05) decrease in plasma testosterone concentration was detected in boars over age. Results suggest that mid-gestational stress of gestating sows does not affect the testosterone concentration or sexual behavior of boar off-spring.     

Early sexual maturity in local boars of Northeastern India: age-related changes in testicular growth, epididymal sperm characteristics and peripheral testosterone levels

The present study reports the age related changes in the peripheral testosterone levels, testicular and epididymal growth and development and cauda epididymal spermiogram in local pigs of Northeastern India, which attain sexual maturity around 3 months of age. Local boars (n = 20) were castrated at monthly intervals from 2 to 6 months of age (4 boars per month) to study the testicular growth and development and the epididymal spermiogram. Blood samples, collected from local boars (n = 6) at monthly intervals from 2 to 6 months of age, were analyzed for testosterone levels by radioimmunoassay. Compared to Hampshire boars, significantly (P < 0.05) high testosterone levels were observed in the local boars as early as 2 months of age. The mean (± SEM) level of testosterone in the local boars at 2, 3 and 4 months of age was 11.89 ± 1.52, 20.45 ± 1.33 and 20.38 ± 2.0 ngml⁻¹, respectively. Though there was consistently significant (P < 0.05) difference in the body weight between Hampshire and local pigs, the same was not observed in case of testicular weight except at 3 and 6 months of age. In line with the above observation, the testis:body weight ratio (gram testis per kg body weight) was significantly (P < 0.05) higher in the local boars compared to the Hampshire boars at any time of observation, which ranged from 0.8 to 1.0 in case of Hampshire and from 2.3 to 3.0 in local boars. The sperm concentration in the cauda epididymal fluid of local boars at 2, 3 and 6 months of age was 2255 ± 186.6, 3685 ± 103.8 and 4325 ± 146.2 million/ml, respectively and the sperm motility, viability and total abnormality was 73.3, 75.2 and 6.2%, respectively at 3 months of age. Taken together, the testosterone level, testicular growth and development and epididymal spermiogram indicate the trait of early sexual maturity in the local pigs as compared to Hampshire.     

Variation in testosterone production among boards and its relationship to sexual interest and breeding performance

Variation in ability to produce testosterone in response to both GnRH and ACTH administration and quatitative relationships between GnRH-stimulated testosterone levels, ACTH-stimulated testosterone levels, sexual interest and breeding performance were assessed in a group of 31 Duroc boars (115.4 +/- 2.5 kg body weight and 212.2 +/- 3.0 days of age). Mean area beneath the testosterone response curve increased (P<0.01) after GnRH and ACTH but the magnitude of response was variable among boars. Post-GnRH testosterone area varied from 7.44 to 50.86 ng/ml X h with a CV = 52.41% while post-ACTH testosterone area varied from 4.99 to 28.78 ng/ml X h with a CV = 45.46%. Mean sexual interest and mean breeding performance scores were correlated (r = 0.67, P<0.01); however, correlations of either variable with testosterone areas were low and nonsignificant. These results indicate that the testosterone-producing ability of boars of similar age and breeding is highly variable and suggest that peripheral testosterone concentrations may not be good indicators of either libido or breeding performance.     

Variability in relationships between semen quality and estimates of in vivo and in vitro fertility in boars

The present experiment was designed to characterize relationships between common semen quality and fertility estimates for three boars known to differ in farrowing rate, number of pigs born alive, and monospermic penetration rate. The approach chosen to accomplish this was to monitor semen quality from these boars and use their semen alternately for either artificial insemination or in vitro fertilization for 40 weeks. This strategy relied on the variability in semen quality parameters that normally occurs in an individual boar over time. When comparisons were made among boars, farrowing rates, numbers of pigs born alive, and monospermic penetration rates were significantly different, but progressive motility, normal head and tail morphology, and acrosome morphology were not. However, when comparisons were made among ejaculates within individual boars, there were significant effects of semen quality on both in vivo and in vitro fertility. For boar 3495, the proportion of spermatozoa exhibiting progressive motility and distribution of spermatozoa in a percoll gradient had a positive linear effect on number born alive and monospermic penetration rate, respectively. For boar 2901, quadratic equations best described changes in litter size as a function of progressive motility and normal acrosomes. In addition, monospermic penetration rate increased linearly as normal acrosomes and the proportion of spermatozoa recovered from a percoll gradient increased. For boar 4291, the relationship between progressive motility and number born alive and between normal acrosomes and number of pigs born alive were also quadratic. However, a significant linear relationship was present only between normal acrosomes and monospermic penetration rate. These results demonstrate that simply relying on the means of common semen quality estimates from some boars has limited value in terms of being used as a prospective indicator of their in vivo or in vitro fertility. In contrast, characterization of relationships between semen quality and fertility estimates is useful for estimating differences in the fertility of ejaculates from individual boars. However, both quantitative and qualitative differences in these relationships among boars are present and a given semen quality estimate that is a good predictor of in vivo or in vitro fertilization for one boar, may not be applicable for others.     

Growth,testicular and endocrine function of boars exposed to 8, 16 or 24 hours of light daily before puberty

Sixty Yorkshire boars were used in two replicates to evaluate the influence of duration of photoperiod on growth, testicular and reproductive endocrine function. Boars born in May and July were exposed to either 8, 16 or 24 h of fluorescent light (200 1x) daily between 2 and 8 months of age. Body weights were obtained at 2-week intervals. At 4, 6 and 8 months of age, venous cannulae were placed in five boars from each treatment and samples of serum were collected to evaluate concentrations of luteinizing hormone (LH) and testosterone. Samples of boars from each treatment were castrated at 6 and 8 months of age, and weights and sperm content of the testes and epididymides were determined. Body weights of boars were similar for all treatments throughout the experiment. Duration of photoperiod did not affect weights of the testes or epididymides or total numbers of sperm within these tissues at either 6 or 8 months of age. Testicular and epididymidal weights and epididymidal sperm numbers increased between 6 and 8 months of age. Concentrations of luteinizing hormone (LH) and testosterone in serum at 4, 6 or 8 months of age were not affected by duration of photoperiod. These results indicate that growth rate and testicular development of prepubertal Yorkshire boars born in May and July are not influenced by duration of photoperiod.     

Effects of nutritional supplements on testicular size and the secretion of LH and testosterone in Merino and Booroola rams

Six Booroola and six Merino rams were fed either a diet which maintained constant live weight or the same diet plus a supplement of high protein lupin grain for 15 weeks, and changes in live weight and testicular volume were measured. Serial blood samples taken for 24 h before the start and 9 weeks after the treatment began were assayed for plasma LH and testosterone and the resulting profiles were analysed for pulses of both hormones. Five weeks later, the animals were given two intravenous injections of 1 μg gonadotrophin-releasing hormone (GnRH) 1 h apart in order to measure pituitary gland responsiveness. A further week later the animals were injected intravenously with 500 μg human chorionic gonadotrophin (hCG) and the levels of testosterone were measured in samples taken after 1.5 h to estimate the testicular responsiveness.The nutritional supplement stimulated testicular growth in both genotypes, so that at the end of the treatment period the testes had increased significantly (P<0.01) in volume by 66% in the Merinos and by 63% in the Booroolas. The live weights also increased, but by relatively less (34% and 43% for supplemented Merinos and Booroolas). The rates of increase in both testicular size and live weight were similar for the two breeds. There were no significant effects of diet on the tonic secretion of LH or testosterone, or on responsiveness to GnRH or hCG.The intervals between LH pulses were significantly shorter (P<0.05) in Booroola rams than in Merino rams both before and after treatment (5.8 h vs. 11.6 h before treatment). The breed differences in LH secretion were mimicked by the testosterone profiles. In the Booroolas, five of the twelve LH profiles contained groups consisting of two to four individually identifiable pulses, each of which elicited a separate pulse of testosterone. A pulse group was observed in only one profile from the Merinos (P=0.06). There were no significant differences between the genotypes in any other parameter of LH or testosterone secretion, or in their responsiveness to GnRH or hCG.It was concluded that (i) nutritional supplements will stimulate testicular growth in both Merino rams and Booroola rams; (ii) the increase in testicular size does not appear to involve an increase in the responsiveness of the testis to LH; and (iii) there are both qualitative and quantitative differences between the genotypes in the patterns of secretion of LH and testosterone which may be associated with the differences in their fecundity.     

Changes in the testicular binding of luteinizing hormone and plasma testosterone concentrations in the Djungarian hamster subjected to different photoperiods and temperatures and effects of long-term testosterone treatment on the binding

The effects of artificial photoperiod, temperature, and long-term testosterone treatment on testicular luteinizing hormone (LH) binding were studied in adult male Djungarian hamsters. In hamsters transferred to long-day (LD; 16 hr light, 8 hr dark) photoperiod 8 weeks after adaptation in short-day (SD; 8 hr light, 16 hr dark) photoperiod of 25 degrees C, testicular growth was associated with an increase in the total LH binding per two testes and a decrease in LH binding per unit testicular weight. Plasma testosterone levels reached a peak 47 days after transfer to LD and tended to decrease thereafter, while the testes continued growing. In contrast, when hamsters reared under LD conditions at 25 degrees C for 12 weeks were transferred to SD, testicular regression was associated with a decrease in plasma testosterone and the total LH binding per two testes and an increase in LH binding per unit testicular weight. A significant decrease in LH binding per unit weight compared to SD controls was observed in those hamsters exposed to SD with continuous testosterone treatment. The testosterone treatment tended to induce decrease in the total LH binding. Scatchard plot analyses of the binding suggested that changes in LH binding were due to changes in the number of binding sites. When sexually mature male hamsters were subjected for 8 weeks to two different ambient temperatures (7 degrees C and 25 degrees C) and photoperiods (LD and SD), the difference between the two temperature groups was statistically not significant regarding the weights of testes, epididymides, and prostates; plasma testosterone levels; and LH binding in either LD or SD group. These results suggest that photoperiod is a more important environmental factor than temperature for the regulation of testicular activity and LH receptors and that testosterone reduces the number of LH receptors per unit testicular weight in adult male Djungarian hamsters.     

Effects of immunization against GnRH on gonadotropins, the GH-IGF-I-axis and metabolic parameters in barrows

Surgically castrated male piglets (barrows) reveal an increase in LH and a decrease in GH compared to untreated boars. Boars that were castrated by immunization against gonadotropin releasing hormone (GnRH) have decreased LH but maintain GH. The difference in GH levels between barrows and immunological castrated boars cannot be explained by testicular steroids because they are low in surgical and immunocastrated boars as well. Therefore, differences in GH concentrations might be due to an interaction between GnRH and growth hormone releasing hormone (GRH) in the hypothalamus or the pituitary. This hypothesis was tested with twelve male piglets that had been castrated within 1 week postnatally and fitted with indwelling cephalic vein catheters at 17 weeks of age. They were split into a control group and an immunized group (each n = 6). Vaccination with Improvac® was performed at 18 and 22 weeks of age. Specific radioimmunoassays were used for hormone determinations (GH, LH, FSH, testosterone and IGF-I). Additionally, metabolic responses were evaluated by measuring analytical parameters that characterize protein synthesis and breakdown, and body fat content. The second vaccination led to a rapid decrease of LH below the limit of detection whereas FSH decreased more slowly, over a period of 5 weeks, from 2.2 to 0.5 ng/ml. This level of FSH, which corresponds to boar-specific concentrations, was maintained thereafter. GH decreased with increasing age but was not influenced by vaccination and remained at a low concentration typical for barrows. Similarly, IGF-I was not altered by vaccination. Consequently, metabolic status was not changed by immunization. It is concluded that the difference in GH levels between surgical and immunocastrated boars is not explained by an interaction between GnRH and GRH.     

Effect of vaccination against gonadotropin-releasing factor (GnRF) with Bopriva® in the prepubertal bull calf

The aim of this study was to evaluate the effect of immunization against gonadotropin-releasing factor (GnRF) with Bopriva(?) (Pfizer Animal Health, Parkville, Australia) in prepubertal bull calves. For the study, 6 calves were vaccinated at the age of 3 and 6 weeks with 1 mL Bopriva(?), and 6 animals served as matched controls. Concentrations of GnRF antibodies, testosterone and LH were determined in serum samples out to 30 weeks after the first immunization. Body weight and scrotal circumference were measured for 59 weeks. At slaughter, 65 weeks after the first immunization, the quality of epididymal sperm was evaluated. The results showed that vaccination against GnRF influenced (P<0.05) anti-GnRF titer, LH and testosterone concentrations as well as scrotal circumference. Antibody titers significantly (P<0.05) increased after the booster vaccination and reached peak values 2 weeks later. Compared to control animals, inhibition (P<0.05) of the prepubertal LH secretion was observed in vaccinated calves at weeks 10 and 12-14 after the first vaccination. In vaccinated calves testosterone concentrations decreased after the booster injection to values below 0.5 ng/mL serum and remained for at least 22 weeks at this low level. Animals vaccinated with Bopriva(?) showed a delay in testes growth and smaller scrotal circumference. Puberty occurred at the age between 46 and 55 weeks in vaccinated and between 38 and 52 weeks in control animals and body weight gain was similar in both groups. All vaccinated bulls attained spermatogenic capacity at slaughter when they were 68 weeks old.     

Ultrastructural study of the boar seminiferous epithelium: changes in cryptorchidism

The present study compares the ultrastructural features of Sertoli cells and germ cells between scrotal testes of healthy boars and abdominal testes of unilateral and bilateral cryptorchid boars. In healthy boars, spermatogonia are flat cells lying in close association with the basal lamina. As differentiation progresses, spermatogonia acquire an oval profile and lose their contact with the basal lamina. Spermatocytes are round cells moving from the basal compartment of the seminiferous epithelium to the luminal compartment. Spermatids exhibit complex morphological changes leading to the formation of spermatozoa. Sertoli cells extend from the basal lamina to the tubular lumen. The nucleus encloses fine euchromatin and one or two nucleoli; the nuclear envelope has a few deep infoldings. The lateral cell membranes form junctional specializations that constitute the blood-testis barrier. The cytoplasm encloses smooth endoplasmic reticulum, vesicles, aggregates, and scattered mitochondria. The seminiferous epithelium of abdominal testes from unilateral and bilateral cryptorchid boars contains few spermatogonia with an abnormal appearance; the alteration in germ cell number is more severe in the bilateral disease. In unilateral cryptorchid boars, spermatogonia appear as either large pyramidal cells or roundish cells; in bilateral cryptorchid boars, spermatogonia show roundish profiles and degenerative patterns. Abdominal testes of both unilateral and bilateral cryptorchid boars are constituted by immature Sertoli cells that show abnormal cytoplasmic content, defective development of the blood-testis barrier, and atypical nuclear appearance; in bilateral cryptorchid boars, immature Sertoli cells exhibit degenerative signs. At postpubertal age, unilateral and bilateral cryptorchidism induce total arrest of spermatogenesis at spermatogonial stage as a result of an abnormal differentiation of the Sertoli cells. Moreover, the degeneration of abdominal testes initiates earlier in bilateral cryptorchidism than in unilateral cryptorchidism.