Pattern of gonadotropin secretion and ultrasonographic evaluation of developmental changes in the testis of early and late maturing bull calves

This was a study that retrospectively analyzed serum gonadotropin secretion and the ultrasonographic appearance of the testis during development in prepubertal bull calves to determine whether there were differences between early and late maturing bulls. Blood samples were taken every other week from 2 wk of age until puberty. Samples were also taken at 12 minute intervals for 12 hours at 4, 10, 20, 25, 30, 35, 40 and 45 wk of age. The GnRH treatment was administered 10 hours after the start of each period of frequent blood sampling. Bull calves fell into two distinctive groups, with one group maturing between 36.6 and 44.2 wk (n = 12) and the other between 46.4 and 48.9 wk of age (n = 8). In samples taken every other week mean serum LH concentrations were greater in early maturing bulls than in late maturing bulls at 12, 14 and 16 wk of age (P<0.05). In blood samples taken every 12 minutes for 10 hours early maturing bull calves had higher mean serum LH concentrations at 4 and 10 wk of age (P<0.05) and higher LH pulse frequency at 10 and 20 wk of age (P<0.05). Mean serum LH concentrations at 4, 10 and 40 wk of age and LH pulse frequency at 10 and 20 wk of age were negatively correlated with age at puberty in bull calves. Mean pixel units of the right and left testis were higher from 34 to 40 wk of age in early maturing than in late maturing animals (P<0.05). It seems possible that hormone measurements and ultrasonographic characteristics of the testes could be developed into powerful tools for studies on the regulation of reproductive development and may aid in the prediction of reproductive potential.     

Effects of season of birth on the prepubertal pattern of gonadotropin secretion and age at puberty in beef heifers

There is an early transient rise in gonadotropin secretion in spring-born prepubertal heifers and there is an indication that this pattern is different in autumn-born heifers. The effect of season of birth on age and weight at puberty is equivocal. This study was designed to compare the temporal patterns of LH and FSH secretion between spring- and autumn-born heifers and to determine the effects of season of birth on age and weight at puberty. Blood samples from 2 groups of heifer calves born in spring (last week of March, n = 5) or autumn (last week of October, n = 5) were collected every other week from birth to puberty and every 15 min for 10 h at 6, 12, 18, 24 and 32 wk of age. Timing of puberty was determined by measuring progesterone in plasma samples collected every 2 to 3 d starting at 42 wk of age. Age and weight at onset of puberty did not differ between the 2 groups of heifers (P > 0.05); however, the autumn-born heifers tended to mature in a wider range of ages and weights. Based on the 10-h sampling periods, mean serum concentrations of LH and LH pulse frequency and amplitude were higher in spring-born heifers at 18 wk of age than in autumn-born heifers (P < 0.05). In spring-born heifers, LH pulse frequency increased over time to 32 wk of age, and LH pulse amplitude was higher at 12 and 18 wk than at 32 wk of age (P < 0.05). Autumn-born heifers had higher LH pulse frequency at 6 wk and showed a decrease in mean concentrations of LH at 12 and 18 wk of age (P < 0.05). The FSH pulse frequency of spring-born heifers was higher at 12 wk of age than in autumn-born heifers (P < 0.05), FSH pulse amplitude in autumn-born heifers decreased from 6 to 32 wk of age. It was concluded that although the mean age and weight at puberty did not differ between spring- and autumn-born heifers, the range in age and weight at puberty was wider in the autumn-born heifers. The patterns of LH secretion differed between spring- and autumn-born prepubertal heifers, with spring-born calves exhibiting an early rise in LH secretion, while mean serum concentrations of LH decreased during this period in autumn-born heifers.     

The relationship between secretory patterns of gonadotrophic hormones and the attainment of puberty in bull and heifer calves born early or late during the spring calving season

It was suggested that an early increase in gonadotrophin secretion in calves aged between 6 and 24 weeks might be critical for initiating developmental changes culminating in puberty. An early rise in luteinizing hormone (LH) release appears to be caused by an increase in LH pulse frequency in bull calves and by an increase in LH pulse amplitude in heifer calves. Previously we have found differences in the characteristics of the LH rise between prepubertal beef calves born in spring or fall; however, age at puberty was not affected by season of birth. Here we report the LH/FSH secretory patterns in prepubertal bull and heifer calves (Hereford x Charolais), born in March or April, respectively (i.e., early or late during the spring calving season; six animals of each sex born at each time). The bull calves of both groups reached puberty (defined as an attainment of scrotal circumference of >or=28 cm) at 43.2+/-1.3 weeks of age (P>0.05). Age at puberty for March- and April-born heifer calves (defined as the age at which serum progesterone concentrations first exceeded 0.4 ng/ml) averaged 56.0+/-1.4 weeks (P>0.05). Based on blood samples taken weekly from birth to 26 weeks of age, and then every other week until puberty, bull calves born in March exceeded April-born bull calves in mean serum LH concentrations at 6, 10 and 12 weeks of age (P<0.05). Mean FSH concentrations were greater (P<0.05) in March-born compared to April-born bull calves from 34 to 32 weeks before puberty. Mean serum LH (at 40, 42 and 56 weeks) and FSH concentrations (at 2, 10, 20, 22-26, 30 and 56 weeks of age) were greater (P<0.05) in heifer calves born in April than March. On the basis of frequent blood sampling (every 12 min for 10 h), heifer calves born in April exceeded March-born animals in mean LH and FSH concentrations, at 5 and 25 weeks, and LH pulse frequency, at 5, 10 and 25 weeks of age (P<0.05). None of the parameters of LH secretion (i.e., mean concentrations of LH, LH pulse frequency and amplitude based on frequent blood collection) differed between March- and April-born bull calves in this study (P>0.05). In summary, March-born bull calves had greater mean serum LH and FSH concentrations prior to 24 weeks of age than April-born calves. April-born heifer calves had greater mean serum concentrations of LH and FSH but this difference was not confined to the early postnatal period. Although there were significant differences in absolute amounts of LH secreted, there were no differences in the frequency of LH secretory pulses amongst March- and April-born bull calves and no differences in LH pulse amplitude in heifer calves born in March or April. As these particular parameters of LH secretion, as well as age at puberty, are not affected by the time or season of birth, they may be primary hormonal cues governing sexual development in bulls and heifers, respectively.     

Plasma LH,FSH, testosterone,and age at puberty in ram lambs actively immunized against an inhibin alpha-subunit peptide

Active immunization against inhibin has been shown to advance puberty and increase ovulation rate in ewe lambs; but in ram lambs, effects on puberty and sperm production are equivocal. The objective of the present study was to determine whether active immunization against an inhibin alpha-subunit peptide advances the onset of puberty in ram lambs. St. Croix hair sheep ram lambs were assigned to inhibin-immunized (n = 7) and control (n = 8) treatment groups. Lambs in the inhibin-immunized group were immunized against a synthetic peptide-carrier protein conjugate, alpha-(1-25)-human alpha-globulin (halpha-G), and control lambs were immunized against halpha-G. Lambs were immunized at 3, 7, 13, 19, 25, 31, and 37 weeks of age. On the day of immunization a blood sample was collected and lambs were weighed. Another blood sample was collected 1 week following each immunization. At 20 weeks of age additional blood samples were collected at 20 min intervals for 8h. Beginning at 20 weeks of age and at weekly intervals thereafter, scrotal circumference (SC) was measured and semen was collected using electroejaculation. A subsequent ejaculate was collected 1 week following onset of puberty, which was defined as the week of age when an ejaculate first contained > or =50 x 10(6) sperm cells. In control lambs, plasma alpha-(1-25)-antibody (Ab) was nondetectable. In inhibin-immunized lambs, alpha-(1-25)-Ab titer increased from 7 to 25 weeks of age and then plateaued at a level that varied (P<0.001) among animals. Body weight and SC of control and inhibin-immunized lambs were similar at the onset of puberty. At pubertal onset inhibin-immunized lambs were older than control lambs (31.9+/-0.5 vs. 29.5+/-0.7 weeks of age, P<0.05). Plasma FSH concentrations were similar in control and inhibin-immunized lambs from 3 to 38 weeks of age. Plasma LH levels were lower (P<0.01) in inhibin-immunized than control lambs. During the 8-h blood sampling period at 20 weeks of age, LH and testosterone concentrations were lower (P<0.05) in inhibin-immunized than control ram lambs, and the LH pulse frequency was similar in the two groups of animals. The decreased LH secretion is consistent with the immunoneutralization of a putative inhibin alpha-subunit-related peptide that stimulates LH secretion in ram lambs. Present findings show that active immunization against an inhibin alpha-peptide delays rather than advances puberty in ram lambs.     

Production and endocrine role of inhibin during the early development of bull calves.

This study investigated the ontogeny of control of FSH secretion by inhibin during early prepubertal development of bulls by 1) measurements of circulating levels of inhibin and FSH from 1 to 13 wk of age, and 2) immunoneutralization of endogenous inhibin at 7, 21, 60, and 120 days of age. In addition, production and localization of inhibin in testes were examined by immunohistochemistry and Western blots at 7, 21, 60, and 120 days of age. Plasma immunoreactive inhibin levels were relatively low between 1 and 3 wk of age and then showed a tendency to rise (P < 0.1) from 4 wk of age. Circulating concentrations of FSH were low during 3 wk after birth and increased at 5 wk, remained high (P < 0.05) until 16 wk of age. Treatment with inhibin antiserum resulted in a significant (P < 0.05) increase in plasma FSH at 7, 21, 60, and 120 days of age compared to those following injection of control serum; however, the magnitude of the FSH rise after inhibin immunization was greater as bulls aged. There were no significant changes in plasma LH after inhibin immunization. An intense staining of inhibin alpha subunits was found in Sertoli cells within the solid seminiferous cords from 7 to 120 days of age, while no specific immune reaction was found in interstitial cells. Western blot analysis of testicular homogenates isolated from bulls 7-120 days of age revealed presence of a 28.5-kDa molecule that cross-reacted with inhibin alpha subunit and beta(B) subunit-specific antibodies. In this study, before 13 wk of age in bull calves, there was no inverse relationship between plasma concentrations of immunoreactive inhibin and FSH. However, the present immunization study clearly indicates that inhibin participates in the regulation of FSH secretion from infancy to early prepubertal stage, although the endocrine significance of inhibin becomes greater in older bulls. The results also indicate that the major production site of inhibin in the testis is Sertoli cells and that these cells produce inhibin that exerts a negative feedback effect on FSH secretion from early stages of development.     

Effect of precocious collection on semen output and quality in young Holstein bulls

Semen production units compete heavily with each other, so they tend to select and collect bulls at the earliest possible age, even before puberty, in order to reduce the interval between generations. This study is a retrospective analysis of the effect of precocious collection on semen quality in Holstein bulls. The semen parameters of early- and late-maturing bulls collected before and after 410 days of age, respectively, were compared over two periods, 1991-1995 and 1997-1999. These periods were defined in relation to the collection rhythms (three collections of two ejaculates at 15 days interval before 1996 and adaptation of the collection rhythms to individual physiological capacity after 1996) and the collection conditions. The effects of age, precocious collection and the interaction between age and precocious collection on mean semen parameters (volume of the ejaculate, sperm motility, percent of motile spermatozoa per ejaculate, total sperm concentration and mobile sperm concentration) measured on collections 1-6 (n = 358 for 1991-1995 and n = 121 for 1997-1999), 7-12 (n = 255 for 1991-1995 and n = 80 for 1997-1999) and 13-18 (n = 92 for 1991-1995 and n = 36 for 1997-1999) were studied by covariance analysis. The semen quality of bulls collected at the early age differed from that of bulls collected after 410 days of age for the first period when the collection rhythm was intense. No effect of precocious collection was evidenced for the second period, suggesting the importance of individual adaptation of the collection rhythm to sexual maturation in young bulls. Early collections at a semen production unit reduced the time needed to obtain the number of insemination straws required for the progeny-testing program by 40 days. Early sperm collection is, thus, of economic and technical interest in well managed semen production units.     

Depression of follicle stimulating hormone in bulls injected with follicular fluid

Eight bulls were divided into two groups and injected with either charcoal-extracted steer blood serum or charcoal-extracted bovine follicular fluid (bFF). Ten-milliliter injections were given subcutaneous every 12 h for 4 wk. Jugular blood collected before, during and after the injection period was analyzed for follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by radioimmunoassay. All bulls were exposed to restrained, estrual heifers for 15 min every 2 wk for 16 wk starting 4 wk before the first injection. The number of mounts and services by each bull was recorded. Semen was collected with an artificial vagina and evaluated on alternate weeks during the same period. The concentration of FSH in serum decreased (P < 0.05) by 12 h after the first injection and remained 61% lower than that of serum-injected bulls during the injection period. The concentration of FSH increased (P < 0.05) by 3 d after the last injection. Injections of bFF did not affect the concentration of LH in serum. Bovine follicular fluid injections significantly depressed FSH; however, libido, serving capacity, and semen characteristics were unchanged.     

Effects of treatment with GnRH from 4 to 8 weeks of age on the attainment of sexual maturity in bull calves

In bull calves an early transient increase in circulating concentrations of LH occurs between 6 and 20 weeks of age. This has been shown to influence reproductive development and performance later in life. In an attempt to hasten the onset of sexual maturity, bull calves (Hereford x Charolais) were treated (im) with 120 ng/kg of GnRH (n=6) twice every day from 4 to 8 weeks of age; control calves received saline (n=6). Injection of GnRH resulted in an LH pulse in all animals. GnRH treated bulls displayed more rapid testicular growth rates between 22 and 44 weeks of age. Sexual maturity (SC>or=28 cm) was achieved earlier in GnRH treated bulls compared to saline treated bulls (41.7+/-2.22 and 47.0+/-0.45 weeks of age, respectively) and this was confirmed by age of sexual maturity based on ejaculate characteristics (>50 million spermatozoa, >10% motility; 45.0+/-0.86 and 49.0+/-1.13 weeks of age for GnRH and control treated bull calves, respectively; P<0.05). We concluded that treatment with GnRH, twice daily, from 4 to 8 weeks of age, prior to the endogenous early increase in plasma LH concentrations, could increase in plasma LH concentrations, advance testicular development and reduce age at puberty in beef bull calves. This may provide the basis for a simple regimen to hasten sexual development in the bull calf.     

Effect of long-term immunization against inhibin on sperm output in bulls.

To determine the effect of neutralization of inhibin on sperm output, 12 Holstein bulls were paired by birth date and weight on Day 1 of age. Each bull was actively immunized against bovine inhibin alpha1-26 gly-tyr (bINH) conjugated to human alpha globulin (HAG, n = 6 bulls) or HAG alone (controls, n = 6) at 60 days of age; booster immunizations were administered at 90, 104, 124, 270, and 395 days of age. Body weights and scrotal circumferences were measured at the time of primary immunization and at 10 days after each booster. In addition, jugular blood was obtained at 60, 70, 100, 114, 134, 280, and 405 days of age, during the 3-wk sperm collection period, and during a 6-h blood-sampling period after sperm collection to determine bINH antibody titer and concentrations of FSH, LH, testosterone, and estradiol. Beginning at 405 days of age, sperm output was measured 3 days/wk for 3 wk with two successive ejaculates collected each day for a total of 18 ejaculates per bull. During Days 60-405 of age, the increase in titer of bINH antibodies, scrotal circumference, and serum concentration of FSH was greater (p < 0.01) for the bINH-immunized compared with control bulls. There were significant (p < 0.01) pair x treatment interactions for sperm output and serum FSH and LH concentrations. Specifically, bINH-immunized bulls for four of the six pairs had nearly 50% greater serum FSH concentrations and sperm output. For the remaining two pairs, sperm output was lower and FSH was either lower or only marginally higher in the bINH-immunized bulls compared with controls. Also, the control bulls for the two remaining pairs produced more sperm than all but one bINH-immunized bull, and had markedly higher serum LH concentrations than all other bulls. To summarize, enhancement of sperm output after immunization against inhibin depends on the subsequent increment in FSH concentrations. We conclude that inhibin suppresses spermatogenesis. Thus, methods to immunoneutralize inhibin may have merit as a therapeutic route to enhance sperm production in reproductively maturing bulls.     

Changes in serum luteinizing hormone (LH) concentrations in response to luteinizing hormone releasing hormone (LHRH) in bull calves that attained puberty early or late

The objectives of this study were to determine if the response to luteinizing hormone releasing hormone (LHRH) could be used to select bull calves capable of early sexual maturation and to establish the optimum route and dose of LHRH. In Trial 1, at 4, 10 and 20 week of age, 20 calves were treated iv with 2 microg/kg body weight of LHRH 1 and 5h after commencing a 9-h period of blood sampling. Bulls were separated into early and late maturing (n=10), based on age at puberty (scrotal circumference (SC) of >or=28 cm). At 4 and 20 week of age, peak serum LH concentrations and area under the LH response curve in response to LHRH were lower (P<0.05) in early- versus late-maturing bulls. In Trial 2, calves at 20 week of age were given LHRH as follows: 2 microg/kg body weight iv (n=6), im (n=6) or sc (n=6); 5 microg/kg im (n=6), or ischio-rectally (ir, n=6) or sc (n=6); and 10 microg/kg im (n=6) or sc (n=6). Serum LH concentrations were at a plateau from 30 to 165 min after treatment with 5 microg/kg of LHRH (im or ir; P>0.05). We concluded that the LH responses to LHRH in calves at 4 and 20 week of age could facilitate the development of a simple test (one blood sample prior to treatment with LHRH and a second during the period of sustained response to LHRH) to select early-maturing bulls.     

Adrenal-pituitary-gonadal relationships and ejaculate characteristics in captive leopards (Panthera pardus kotiya) isolated on the island of Sri Lanka

In Study 1, semen was collected using a standardized electroejaculation procedure. Males (N = 8) produced ejaculates with a high incidence of sperm abnormalities (77 +/- 3.3%). After electroejaculation under anaesthesia, serum cortisol concentrations increased (P less than 0.05), while testosterone concentrations decreased (P less than 0.05) and LH and FSH concentrations were unchanged (P less than 0.05) over a 2-h bleeding period. In Study 2, male and female leopards were bled at 5-min intervals for 3 h and given (i.v.): (1) saline (N = 2/sex); (2) GnRH (1 microgram/kg body weight) 30 min after the onset of sampling (N = 5/sex); or (3) ACTH (250 micrograms) at 30 min followed by GnRH 1 h later (N = 5/sex). Basal concentrations of serum LH, FSH and cortisol were comparable (P greater than 0.05) between male and female leopards. After GnRH, peak LH concentrations were 2-fold greater (P less than 0.05) in males than females while FSH responses were similar. In males, testosterone concentrations increased 2-3-fold following GnRH. After ACTH, serum cortisol concentrations doubled within 15 min in both sexes. Administration of ACTH 1 h before GnRH did not affect GnRH-induced LH or FSH release (P greater than 0.05); however, testosterone secretion was only 30% of that observed after GnRH alone (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Patterns of puberal development in Sahiwal and Brahman cross bulls in tropical Australia II. LH and testosterone concentrations before and after GnRH

Plasma LH and testosterone (T) concentrations were measured before (basal) and two hours after (peak) GnRH stimulation in 52 Bos indicus strain bulls between one and two years of age. The animals comprised 13 1 2 Brahman, 20 3 4 Brahman, 8 1 2 Sahiwal and 11 3 4 Sahiwal cross bulls and samples were collected at approximately seven week intervals. Basal- and peak-T concentrations increased between one and two years of age, and basal LH concentrations decreased; no changes in peak LH were noted over time. Peak-T concentrations were significantly correlated with scrotal circumference (SC), sperm per ejaculate and seminal fructose. Significant genotype differences were noted, Sahiwal cross bulls had higher peak-T concentrations at puberty than Brahman cross bulls.     

Electroejaculation, semen characteristics and serum testosterone concentrations of free-ranging african elephants (Loxodonta africana)

A regimented electroejaculation protocol (120 electrical stimulations; 10-30 V) was used to collect semen and characterize ejaculate quality from 9 adult, free-ranging African elephants under anaesthesia. Eight of the 9 ejaculates contained high concentrations of progressively motile spermatozoa. The overall mean ejaculate volume, sperm concentration/ml ejaculate, sperm motility, sperm status and ejaculate pH were 93.3 ml, 2408.6 X 10(6) spermatozoa/ml, 70%, 3.9 and 7.4, respectively. A high percentage (mean 77.5%) of spermatozoa within each ejaculate was morphologically normal. Of the aberrant spermatozoa, 72% had a cytoplasmic droplet defect. When sperm viability was tested in vitro at 37 degrees C, sperm motility rating declined by at least half of the initial assessment within 3.5 h of semen collection. Generally, spermatozoa maintained motility in vitro for less than 6 h. Serum testosterone ranged from 1.4 to 8.2 ng/ml in 4 males evaluated in the morning (07:30-08:00 h). In 4 of the 5 bulls assessed in the afternoon (15:00-18:00 h), testosterone levels were less than 0.9 ng/ml. The remaining bull, evaluated at 16:00 h, had exceptionally high testosterone concentrations (peak 25.6 ng/ml) and a preputial discharge potentially indicative of 'musth'. The present study demonstrates that high quality semen can be collected consistently from the African elephant and that striking differences exist in serum testosterone amongst free-ranging males which may be due, in part, to a diurnal rhythm.     

Effect of age and environmental factors on semen quality, glutathione peroxidase activity and oxidative parameters in simmental bulls

Taking into account that semen quality depends on animal age and climate conditions and that oxidative stress has been reported to be a common cause of infertility, the objective of this study was to monitor indicators of oxidative stress and antioxidant protection during four seasonal periods in service bulls of various age to get better insight into the significance of these factors upon evaluating service bull semen. The research was conducted over a year on 19 Simmental service bulls. Animals were divided into two groups according to age; Group I consisted of younger bulls aged two to four yrs (n = 9), and Group II was comprised of older bulls aged five to ten yrs (n = 10). Semen samples were obtained once in the middle of every seasonal period and blood samples for biochemical analysis were collected by jugular venipuncture immediately after ejaculate collection. The activity of total glutathione peroxidase (T-GSH-Px), selenium-dependent glutathione peroxidase (Se-GSH-Px) and selenium-independent glutathione peroxidase (non-Se-GSH-Px), together with the intensity of lipid peroxidation (thiobarbituric acid reactive substances; TBARS) and oxidative protein damage (protein carbonyl content (PCC)) were measured in seminal plasma. In samples of spermatozoa and blood serum, the activity of Se-GSH-Px and TBARS and PCC concentrations were determined. Older service bulls had significantly higher ejaculate volume in summer in comparison with younger bulls, whereas the number of spermatozoa and progressive motility percentage did not significantly vary with age. Younger animals had lower progressive motility percentage during summer than in spring, with more intensive oxidative processes observed in seminal plasma (TBARS) and spermatozoa (TBARS and PCC). Based on the results presented here, it can be concluded that younger bulls are more sensitive to elevated ambient temperatures during the summer, when intensified prooxidative processes in semen plasma and spermatozoa eventually led to decreased sperm progressive motility with consequential semen quality deterioration.     

Effect of different levels and sources of zinc supplementation on quantitative and qualitative semen attributes and serum testosterone level in crossbred cattle (Bos indicus x Bos taurus) bulls

An experiment was conducted on 16 crossbred bulls (about 2 years of age, 316.2+/-0.77 kg average body weight), divided into groups I, II, III and IV to study the effect of different levels of Zn supplementation from inorganic and organic sources on semen quality. The animals in the first 3 groups were supplemented with 0, 35 and 70 ppm Zn from Zn sulfate, respectively and the animals in-group IV were supplemented with 35 ppm Zn as Zn propionate. Semen collection and evaluation was done in the first month (to assess semen quality at the start of the experiment) and 7th, 8th and 9th month of experimental feeding to evaluate the effect of supplemental Zn on semen attributes. We gave 6 months for Zn feeding, so that 3 sperm cycles of spermatogenesis had passed and the collected semen reflected the complete effect of Zn supplementation. Six ejaculates from each bull were collected and evaluated for semen quantitative (ejaculate volume, sperm concentration and sperm number per ejaculate) and qualitative characteristics (semen pH, mass motility, individual motility, sperm livability percent and abnormal sperm percent, percent intact acrosome, bovine cervical mucus penetration test, hypo-osmotic sperm swelling test) and activity of seminal plasma enzymes i.e., alkaline phosphatase, acid phosphatase, GOT and GPT. Testosterone level in the blood serum of crossbred bulls was also estimated. Mean values of semen quantitative and qualitative characteristics at the start of the experiment were statistically non significant (P > 0.05) in all the crossbred cattle bulls, however, there were statistically significant differences among the bulls of different groups after 6 months of zinc supplementation. Mean ejaculate volume (mL) was 2.37, 4.70, 5.86 and 6.38, respectively in groups I to IV, indicating a statistically significant (P < 0.05) higher semen volume in Zn-supplemented groups as compared to the control group of bulls. Similarly, sperm concentration (million.mL(-1)), live sperm (%) and motility (%) were significantly (P < 0.01) higher in Zn-supplemented groups as compared to the control group. The results of BCMPT and HOSST revealed a significant improvement in sperm functional ability in all the groups supplemented with Zn as compared to the control group. The activity of alkaline and acid phosphatase in seminal plasma was significantly (P < 0.05) higher in the Zn-supplemented groups, whereas GOT and GPT activities in seminal plasma were significantly (P < 0.05) lower in the Zn propionate supplemented group as compared to the control group. Testosterone concentration (ng.mL(-1)) in blood serum was significantly higher in animals of groups III and IV, as compared to control group. It may be concluded that Zn supplementation either in the inorganic or organic form in the diet of crossbred bulls improved the qualitative and quantitative attributes of semen; however, the number of sperm per ejaculate, mass motility and semen fertility test like bovine cervical mucus penetration was significantly higher in bulls given Zn in an organic form (Zn propionate) as compared to an inorganic form (Zn sulfate).     

Sexual development of dairy bulls in the Mexican tropics

Sexual development and pubertal traits were studied in Holstein Frisian (Ho) and Brown Swiss (BS) bulls born and maintained under tropical conditions. Characteristics evaluated every 2 weeks, from 27 to 63 weeks of age, included live weight, scrotal circumference, testicular diameter, semen quality and sexual behavior. Puberty was defined as the age at which a bull first produced an ejaculate containing at least 50 x 106 spermatozoa, with a minimum of 10% progressive motility. Testicular growth was linear in Ho bulls and quadratic in BS bulls. There was no breed difference in age at puberty (Ho, 333 +/- 15.8 days; BS, 311 +/- 10.5 days). However, at puberty, live weight and scrotal circumference tended to be greater in Ho (276 +/- 16.9 kg and 28.4 +/- 1 cm, respectively) than in BS bulls (233 +/- 11.3 kg and 25.9 +/- 0.7 cm, respectively), and testicular diameter was larger for Ho (5.5 +/- 0.24 cm) than for BS bulls (4.8 +/- 0.16 cm). Pooled data for all bulls for semen characteristics at puberty were: volume, 6.3 +/- 0.6 ml; progressive motility, 26.8 +/- 4.4%; sperm concentration, 58.5 +/- 13.9 x 10(6) spermatozoa/ml, and 351.5 +/- 91.2 x 10(6) spermatozoa/ejaculate. These values improved until at least 18 weeks after puberty. Eighty-five percent of bulls mounted heifers by 206 days of age, but only a few bulls had mounts with ejaculation during the study. It was concluded that reproductive development was similar between Ho and BS bulls, but slower than that reported for dairy bulls in temperate areas. Variation in some characteristics, such as scrotal circumference, was observed among bulls within each breed group, which might be of benefit for genetic selection.     

Sexual development in early- and late-maturing Bos indicus and Bos indicus x Bos taurus crossbred bulls in Brazil

Two experiments were conducted to evaluate sexual development in early- and late-maturing Nelore (Bos indicus) and Canchim (3/8 Bos indicus x 5/8 Bos taurus crossbred) bulls and to determine predictors of sexual precocity, and pubertal and maturity status. In Experiment 1, 12 Nelore bulls where examined from 300 to 900 days of age. Puberty was characterized by an ejaculate containing > or =50 million sperm with > or =10% motile sperm, and maturity by an ejaculate containing > or =70% morphologically normal sperm. In Experiment 2, 28 Canchim bulls where examined from 295 to 488 days of age and puberty was characterized by an ejaculate containing > or =30% motile sperm. In both experiments, bulls were classified as early- or late-maturing based on age at puberty. Early-maturing bulls were younger (P < 0.05) than late-maturing bulls at puberty (527 days versus 673 days in Experiment 1 and 360 days versus 461 days in Experiment 2) and at maturity (660 days versus 768 days in Experiment 1). In general, early-maturing bulls were heavier and had greater scrotal circumference (SC), testes, and testicular vascular cone diameter than late-maturing bulls during the experimental period. Scrotal circumference adjusted for 365 days of age was a good predictor of sexual precocity; minimum yearling SC of 19 and 24 cm for Nelore and Canchim bulls, respectively, had the best predictive values. Early-maturing bulls were lighter and had smaller SC at puberty than late-maturing bulls; therefore, sexual precocity was not related to the attainment of a threshold body weight or testicular size earlier, but to lower thresholds in early-maturing bulls. When predictors of pubertal status were evaluated, SC had the best sensitivity/specificity relationship in Nelore bulls, and high sensitivity and specificity in Canchim bulls. When predictors of sexual maturity were evaluated in Nelore bulls, age, weight, and SC had similar sensitivity, specificity, and predictive values. At puberty, approximately 60% of the sperm present in the ejaculate were morphologically defective. Changes in semen quality after puberty in Nelore bulls were characterized by increased motility and proportion of morphologically normal sperm, with a decrease in the proportion of major sperm defects. In conclusion, early-maturing bulls were more developed in the pre-pubertal period and attained puberty at earlier stages of body and testicular development than late-maturing bulls. Yearling SC could be used to select bulls for sexual precocity and SC was the best predictor of pubertal status. Age, weight, and SC were equally good predictors of sexual maturity in B. indicus bulls.     

Gonadotrophin secretion in prepubertal bull calves born in spring and autumn

The reproductive development of bull calves born in spring and autumn was compared. Mean serum LH concentrations in calves born in spring increased from week 4 to week 18 after birth and decreased by week 24. In bull calves born in autumn, mean LH concentrations increased from week 4 to week 8 after birth and remained steady until week 44. LH pulse amplitude was lower in bull calves born in autumn than in calves born in spring until week 24 of age (P < 0.05). There was a negative correlation between LH pulse frequency at week 12 after birth and age at puberty in bull calves, irrespective of season of birth, and LH pulse frequency at week 18 also tended to correlate negatively with age at puberty. Mean serum FSH concentrations, age at puberty, bodyweight, scrotal circumference, testes, prostate and vesicular gland dimensions, and ultrasonographic grey scale (pixel units) were not significantly different between bull calves born in autumn and spring. However, age and body-weight at puberty were more variable for bull calves born in autumn (P < 0.05). In a second study, bull calves born in spring received either a melatonin or sham implant immediately after birth and at weeks 6 and 11 after birth. Implants were removed at week 20. Mean LH concentrations, LH pulse frequency and amplitude, mean FSH concentrations and age at puberty did not differ between the two groups. No significant differences between groups in the growth and pixel units of the reproductive tract were observed by ultrasonography. In conclusion, although there were differences in the pattern of LH secretion in the prepubertal period between bull calves born in autumn and spring, the postnatal changes in gonadotrophin secretion were not disrupted by melatonin treatment in bull calves born in spring. Reproductive tract development did not differ between calves born in spring and autumn but age at puberty was more variable in bull calves born in autumn. LH pulse frequency during the early prepubertal period may be a vital factor in determining the age of bull calves at puberty.     

The effect of nutrition on sexual development of bulls

Most bulls that are managed for sale as yearlings are fed high-energy diets in the post-weaning period to maximize rates of gain in body weight. High-energy diets with adequate protein, vitamins and minerals result in a larger scrotal circumference at 1 y of age, however, part of this increase in size is likely due to scrotal fat. It is unclear whether testis size and spermatogenesis is significantly affected by nutritional intake in the post-weaning period. There are indications of an effect of calfhood nutrition on age at puberty and testis size. Scrotal circumference was smaller in yearling bulls raised by first-parity dams, compared to those raised by older dams. This may have been due to lower milk production by first-parity dams, an in utero effect, or both. The effect of reduced calfhood nutrition may be mediated through gonadotropin secretion. Calves destined to become later maturing bulls with smaller testes had lower amounts of LH secretion during the period of the early gonadotropin rise (8-16 wk of age). Furthermore, augmenting circulating LH concentrations at this time by treating calves with GnRH hastened pubertal development. In addition, FSH treatments in calfhood also increased scrotal circumference and hastened spermatogenesis. In that regard, FSH has been considered a main driver of Sertoli cell proliferation in prepubertal animals. Since Sertoli cell multiplication ceases at 20-25 wk of age in bulls, final testis size in bulls is likely determined in calfhood. Four experiments were done to investigate the effects of calfhood nutrition on pubertal development. These studies confirmed that superior calfhood nutrition augmented gonadotropin secretion (which is probably mediated by metabolic hormones); this resulted in larger testes at 1 y of age and an earlier onset of spermatogenesis.     

Electroejaculation increased vocalization and plasma concentrations of cortisol and progesterone, but not substance P, in beef bulls

Electroejaculation is a reliable method of obtaining a semen sample for a bull breeding soundness examination, but is sometimes regarded as painful. Substance P is a neuropeptide involved in the integration of pain, stress, and anxiety. We hypothesized that substance P is a measure of pain in bulls following electroejaculation. The specific objective was to compare vocalization and plasma concentrations of cortisol, progesterone, and substance P immunoreactivity in bulls following electroejaculation. Nine Angus bulls (501.9 ± 14.3 kg) were used. Blood samples were collected at −60, −30, 0, 2, 10, 20, 30, 45, 60, 75, 90, 120 min relative to treatment. At Time 0, bulls were subject to electroejaculation, rectal probe insertion without electroejaculation, or no manipulation. Treatments were administered contemporaneously to three bulls. Treatments were repeated weekly until each bull had received each treatment in a 3 × 3 Latin square design. More bulls (P = 0.0147) in the electroejaculation group vocalized (5 of 9 bulls; 55.6%) when compared to controls (0 of 9 bulls; 0%). Mean plasma cortisol and progesterone concentration following electroejaculation in bulls were higher (P < 0.05) than concentrations in probed and control bulls through the 45 min sample. However, mean plasma substance P concentration following electroejaculation in bulls (77.2 ± 17.2 pg/mL) was not different (P = 0.6264) from probed (79.1 ± 17.2 pg/mL) or control bulls (93.4 ± 17.2 pg/mL). A significant increase in vocalization and plasma cortisol and progesterone concentrations in bulls following electroejaculation was likely owing to acute stress. However, the lack of a difference in plasma concentrations of substance P after electroejaculation was interpreted as a lack of pain associated with nociception.