Importance of non-olfactory ram stimuli in mediating ram-induced ovulation in the ewe

In Exp. 1, 4 groups of 50 recently weaned ewes were exposed to various degrees of contact with rams for 65 days, followed by exposure to novel rams for 4 days. Ovarian activity in the ewes was determined by laparoscopy on Days 29, 65 and 69 of treatment. There were no treatment differences in the percentage of ewes ovulating on Day 4 whereas by Day 29 more ewes in clear fenceline and full ram contact were ovulating compared to controls (P less than 0.05, P less than 0.001). After 65 days ovarian activity was significant only in those ewes in full contact with rams (P less than 0.001). Between 89 and 95% of ewes remaining anovulatory after 65 days ovulated after 4 days of full contact with novel rams. In Exp. 2, 4 groups of about 30 anovulatory ewes were exposed to various degrees of contact with rams for 5 days. Ovarian activity was assessed before and after treatment by laparoscopy. After 5 days, more ewes were ovulating in response to full ram contact than in any other treatment (P less than 0.05) and more ewes in fenceline contact with rams or with rams plus ewes were ovulating than in the isolated control treatment (P less than 0.01). In Exp. 3, 6 groups of about 40 anovulatory ewes were exposed to face masks with and without rams' wool and/or various degrees of contact with rams for 5 days. More ewes were ovulating after 5 days in the group in full physical contact with rams than in any other group (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of classification levels of ram sexual activity on spring breeding ewes

Ram lambs (7–8 months old) and mature rams (19–20 months old) were used to evaluate the effect of classification levels of male sexual performance on reproductive performance of ewes during spring breeding. In Exp. 1, sexually active ram lambs with high (1.8±0.3; n=5) and low (0.9±0.2; n=5) sexual performance scores (HP and LP; mean±S.E.M.) were used in single sire breeding pens. Ewes (n=305) were stratified by age and assigned to 10 pens for 34 days starting in late March. For Exps. 2 and 3, two replicates were conducted for 2 years with sexually active mature rams in a single sire mating scheme. For Exp. 2, HP rams (n=5) averaged 3.6±0.2 ejaculations and LP rams (n=7) 1.8±0.2 ejaculations for sexual performance scores based on nine, 30 min serving capacity tests (SCT). Polypay ewes (n=152 to 153 per year) were stratified by age and assigned to pens each year for 34–38 days starting in late March for years 1 and 2. For Exp. 3, HP rams (n=6) averaged 3.7±0.1 ejaculations and LP rams (n=10) 2.3±0.1 ejaculations for sexual performance scores based on 18, 30 min SCT. Polypay ewes (n=229 in year 3 and n=244 in year 4) were stratified by age and assigned to pens each year for 34 days starting in late March. In Exp. 1, lambing rates for ewes bred to HP versus LP ram lambs did not differ (65.8 versus 53.0; P=0.20). Prolificacy tended (P=0.06) to be increased by 0.1 lambs in ewes bred by LP ram lambs. Total number of lambs born per ewe present at lambing, and lambing distribution were not altered by HP and LP ram lambs. In Exp. 2, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (58.1 versus 60.1; P=0.78). In Exp. 3, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (74.3 versus 69.0; P=0.35). There was no difference (P>0.10) between years for Exp. 2 or Exp. 3, and mature HP and LP rams did not affect the other reproductive variables monitored. Analyses of the combined data for Exps. 2 and 3 indicated only a year difference (P<0.001) in lambing rates and total lambs born. Present studies indicate that different sexual performance classifications for ram lambs and mature rams did not alter lambing rates or distribution of lambing of Polypays bred in late March to April. These results indicate that HP and LP, sexually active, Polypay rams and ram lambs with average to high quality semen can provide a source of rams for spring breeding Polypays in ambient conditions and that there was no advantage to using HP over sexually active LP ram lambs or rams.     

Effects of recent sexual experience and melatonin treatment of rams on plasma testosterone concentration, sexual behaviour and ability to induce ovulation in seasonally anoestrous ewes

The aim of this study was to determine whether advancing the seasonal changes associated with rams by treatment with exogenous melatonin and allowing the rams previous sexual experience would increase the proportion of anoestrous ewes ovulating in early July. North Country Mule ewes (n = 225) were grouped by live body weight and body condition score and allocated randomly to the following treatments: (i) isolated from rams (control; n = 25); (ii) introduced to rams (treatment 2); (iii) introduced to rams that had mated with ewes during the previous 2 days (treatment 3); (iv) introduced to rams implanted with melatonin (treatment 4); and (v) introduced to rams that were implanted with melatonin and had mated with ewes during the previous 2 days (treatment 5). Treatments 2-5 were replicated (2 x 25 ewes) and two rams were introduced to each replicate group. Introductions began on 4 July and were completed by 11 July. The rams were withdrawn from the ewes after 8 days. Melatonin was administered as a subcutaneous implant (Regulin((R))) on 22 May and again on 20 June. Blood samples were taken from all rams to determine plasma melatonin and testosterone concentrations (19 samples in 6 h). The behaviour of the sheep was videotaped continuously during the first 3 h after the ram was introduced. Ovulation was detected by an increase in plasma progesterone concentrations from < 0.5 ng ml(-1) to > 0.5 ng ml(-1). Mean +/- SE plasma melatonin concentrations were 649.7 +/- 281.4 and 18.3 +/- 2.4 pg ml(-1) in rams with and without melatonin implants, respectively (P < 0.001). Melatonin implants also increased plasma testosterone concentrations from 4.30 +/- 1.88 to 10.10 +/- 1.10 ng ml(-1) (P < 0.01), the libido of the rams and the proportion of ewes that ovulated in response to the rams (43 and 56% (treatments 4 and 5) versus 24% (treatments 2 and 3)). In conclusion, implanting rams with melatonin before introducing them to seasonally anoestrous ewes increases the proportion of ewes that ovulate in response to introduction of a ram, but previous sexual experience of rams appears to have little or no effect.     

Luteinizing hormone release in intact and castrate rams is altered with immunoneutralization of endogenous estradiol

Changes in the dynamics of luteinizing hormone (LH) release in the adult ram following immunoneutralization of endogenous estradiol were investigated. Castrate rams were actively immunized against estradiol-6-bovine serum albumin for 7 months and then their patterns of episodic LH release and LH response to multiple injections of gonadotropin-releasing hormone (GnRH, two 5-micrograms doses given iv 2 h apart) were assessed (April). In comparison with control rams immunized against rabbit gamma globulin, estradiol-immunized rams (antibody titre approximately 1:5000) exhibited more frequent LH releases (11.7 +/- 0.3 vs. 9.3 +/- 0.8 pulses/8 h, P less than 0.05) and a greater LH response to the first GnRH injection (peak delta value 190 +/- 8 vs. 130 +/- 25 ng/mL, P less than 0.01). Estradiol antiserum collected from the castrate rams was used in the passive immunization of intact rams (antibody titre approximately 1:200) for 1 month (beginning mid-July). Although episodic LH release was always similar for control and immunized rams, testosterone levels in the latter group increased approximately 150%. In contrast with the castrate ram response, GnRH treatment (two 5-micrograms doses given iv 80 min apart) produced a "self-priming" effect on LH release in the intact rams, an effect that was dampened with estradiol immunoneutralization. Consequently, peak 2:peak 1 ratios for delta value and 80-min mean incremental increase were much smaller (P less than 0.01) for the immunized rams (approximately 2:1 vs. 4:1 for the control rams).(ABSTRACT TRUNCATED AT 250 WORDS)     

The contribution of the male to ovine embryogenesis in an in vitro embryo production system

The differences in the embryo production potential of four rams used in a commercial embryo transfer program were examined in both in vivo and in vitro embryo production systems. Processing frozen-thawed spermatozoa through Percoll density gradients prior to in vitro insemination eliminated differences in the estimates of sperm viability between the four rams, and yet, differences in embryo production persisted throughout the in vitro culture period. However, there was no effect of ejaculate within ram on embryo production rates. In addition, the timing of the onset of the differences between the rams at each stage of in vitro embryo development were revealed. Ram 2 differed from ram 4 in the proportion of fertilized oocytes at 17 h post-insemination (pi) and by 52 h, ram 3 differed from ram 4 in the proportion of cleaved embryos, and the observed differences between ram 1 and ram 2 in their blastocyst production were initiated prior to activation of the embryonic genome. Once differences in embryo development rates were detected among the four rams, they persisted throughout the in vitro culture period. The reduced in vitro fertilization (IVF) rates from ram 2 compared with the other rams was paralleled in vivo by the significantly lower proportion of embryos recovered from ewes mated to ram 2, and this was further exacerbated by a significantly lower embryo survival rate after transfer. However, the subtle differences observed in the timing of the contribution of each sire to embryo development during in vitro culture were not able to be detected in vivo. However, the higher proportions of transferable quality blastocysts obtained from ewes mated to ram 4 did not result in increased embryo survival throughout the remainder of gestation. Therefore, in this study, the blastocyst production potential for a particular sire, either in vitro or in vivo, does not necessarily reflect the potential for the production of live offspring.     

A note on ram preference by oestrous ewes: Influence of rams’ age and sexual performance

Ewes in oestrus, besides being receptive to rams, develop an active search for them. We performed two experiments to determine if oestrous ewes prefer: (1) high or low sexual-performance rams, and (2) 2-year-old or aged (>8-years-old) rams. In the first experiment, the preference of ewes for rams with high or low sexual performances was compared in 16 tests. In the second experiment, 17 tests were performed comparing ewes’ preference for aged rams (>8-years-old) or young rams (2-years-old). In each test, two rams were tied at equal distances from the point of entrance of the ewe, with a maximum mobility for each ram of 2 m. The first approach to a ram, the quantity of approaches toward each ram (less than one body length), and the total time that the ewe remained near each ram were registered during 5 min. Oestrous ewes did not display any preference according to sexual performance of rams, but spent significantly more time (P = 0.02), and did it in more tests (P = 0.025), with young than with aged rams. Ewes also tended to approach first (P = 0.07), and more times in more tests (P = 0.06), to young than to aged rams. We conclude that ram's sexual performance seems not to influence oestrous ewes preference, and that ewes’ prefer young (2-years-old) rather than aged rams (>8-years-old).     

Serum luteinizing hormone, testosterone, and thyroxine and growth responses of ram lambs fed locoweed (Oxytropis sericea) and treated with vitamin E/selenium

Sixteen ram lambs (5 m.o. old, 45 +/- 1.5 kg) received a control diet (50% concentrate, no locoweed, n = 4), locoweed (20% locoweed for 21 d, n = 4), MUSE (2 mL i.m. of MUSE containing 5 mg selenium and 50 mg vitamin E/mL, n = 4) on Days 21 and 35([Day 0 = first day of trial]), or locoweed + MUSE (n = 4). The rams were maintained in individual pens (3 x 9 m) with free access to feed, water, salt and shade. On Day 7 after initiating locoweed, serum alkaline phosphatase (AP) increased (P < 0.01), and serum thyroxine (T4) decreased (P < 0.01) in locoweed-fed rams. Effects on serum AP and T4 remained constant in rams during the 21 d of locoweed feeding. Treatment with MUSE did not influence (P > 0.10) AP or T4. Locoweed-fed rams had reduced (P < 0.05) intake and body weight for the 2-wk period after locoweed feeding ended. The MUSE regimen or diet had no effect on intake or body weight (P > 0.50). Neither locoweed nor MUSE affected serum LH before or after GnRH administration on Day 22 (P > 0.10). On Day 50, however, area under the LH curve (AUC) was 966 units in locoweed-fed rams and 1,373 units (+/- 154) in controls (P = 0.09). Serum testosterone (T) was reduced in locoweed-fed rams before and after (P < 0.05) GnRH on Day 22. On Day 50, the T AUC was numerically lower (P = 0.14) in locoweed-fed rams (1,252 units) than in controls (1,539 +/- 130 units). Conversely, MUSE treatment resulted in increased (P = 0.02) T AUC on Day 50 (1,148 and 1,643 +/- 130 units in control and MUSE-treated ram lambs, respectively). During the 6-wk period after locoweed feeding, serum immunoglobulin G averaged 14.0 and 18.6 (+/- 1.1) mg/mL in control and locoweed-fed rams (P < 0.01), respectively. Twenty percent dietary locoweed for 21 d exerts adverse effects on feed intake, growth, and reproduction in young ram lambs and MUSE was not effective in reversing these effects.     

Short-term treatment with a controlled internal drug releasing (CIDR) device and FSH to induce fertile estrus and increase prolificacy in anestrous ewes

The objectives were to evaluate, in anestrous ewes, the effectiveness of a CIDR-G device (0.3 g progesterone) administered for 5 d to induce estrus; and FSH (Folltropin; 55 mg NIH-FSH-P1 equivalent) in saline:propylene glycol (1:4) 24 h before insert removal (Day 0), to increase ovulation rate and prolificacy. Ewes of mixed breeding were assigned at random to 3 treatments: control (C; n = 125), 5 d progesterone (P5; n = 257) and 5 d progesterone plus FSH (P5F; n = 271). Intact rams were joined at insert removal and ewes were observed every 24 h for 3 d. On Day 14, the ovulation rates of all ewes detected in estrus in the treated groups were determined using transrectal ultrasonography. Rams were removed on Day 26 to 31. Ewes were examined for pregnancy then, and again 20 to 25 d later to detect ewes that conceived to the second service period. Percentage of ewes marked by rams was higher in progesterone-treated (77%) than in C (20%; P < 0.01), but did not differ between P5 and P5F. The ovulation rate (1.95+/-0.04) did not differ due to FSH. Conception (68%) and pregnancy (52%) rates were higher in progesterone-treated (P < 0.01) than in C (0%) ewes. Estrous response varied quadratically with time after ram introduction, and the conception rate varied quadratically with the time of observation of onset of estrus. Over two service periods more progesterone-treated than C ewes lambed (65 vs 45%; P < 0.01). Lambs born per ewe exposed (0.7+/-0.1, 1.0+/-0.1, and 1.1+/-0.1 for C, P5 and P5F, respectively) was increased by progesterone (P < 0.05). Litter size to the first service period (1.59+/-0.04) and overall (1.54+/-0.03) did not differ among treatment groups. FSH-treated ewes tended to have more lambs (1.67+/-0.1) than did ewes receiving progesterone alone (1.5+/-0.1; P = 0.06) and than did ewes lambing to the second service period (1.5+/-0.1; P = 0.06). In summary, a 5-d progesterone pre-treatment of anestrous ewes induced estrous cycles and increased the pregnancy rates. A single injection of FSH only tended to increase litter size.     

The effect of melatonin implants on the response to the male effect and on the subsequent cyclicity of Rasa Aragonesa ewes implanted in April

Rasa Aragonesa ewes were used to evalutate whether treatment with melatonin implants in spring could modify: (i) the response to the male effect in terms of oestrous behaviour and ovulation rate; and (ii) the maintenance of sexual activity and ovulation rate at medium term, i.e. over the next 306 days. On 12 April, 42 ewes were divided into two groups, with (M; n = 21) or without (C; n = 21) a subcutaneous implant containing 18 mg melatonin. On 17 May (day 0), three aproned rams were introduced to each group to induce a ram effect. Ewes were observed for oestrus daily. The rams were removed 40 days later after which one aproned ram was introduced daily. Oestrous detection continued until 28 February, 306 days after the first male-female contact. The ovulation rate was determined by endoscopy in the first three cycles after ram introduction and in September-October and January. Progesterone was assayed from blood samples taken on 6 May, 10 and from day 0 to day 22 after ram introduction. Luteal activity before ram introduction was seen in 33% (M) and 29 (C)% of the ewes, respectively. Significantly more M ewes showed oestrous behaviour during the first 40 days after ram introduction (M: 100%; C: 62%; P < 0.01). Similar differences were observed for ewes anovulatory at ram introduction (M: 100%, C: 47%; P < 0.01). These differences were maintained over the three oestrous cycles in both groups. Treatment with melatonin implants was without detrimental effect on cyclic functions in the following breeding season, after seasonal anoestrus. Melatonin treatment significantly increased (P < 0.05) the mean ovulation rate of the first (1.62 +/- 0.11 versus 1.31 +/- 0.13), second (1.78 +/- 0.15 versus 1.36 +/- 0.15) and third cycles (M: 1.73 +/- 0.12 versus C: 1.27 +/- 0.14). There was a significant interaction between the effects of cyclicity at day 0 and melatonin treatment on the ovulation rate in the first cycle (P < 0.05). The mean ovulation rates of both groups were similar at the beginning (September) and middle (January) of the subsequent breeding season. Overall, the results confirmed that melatonin implants, combined with the ram effect, improved the reproductive parameters of reduced-seasonality ewes during spring mating, without impairing sexual activity or ovulation rate during the subsequent breeding season.     

Unilateral orchidectomy of ram lambs: acute, chronic and hCG-stimulated androgen levels

1. The objectives of this study were to determine whether compensation of androgen secretion occurred acutely, chronically or after hCG-stimulation in unilaterally orchidectomized (ULO) rams. 2. Testosterone (T) concentrations were not significantly different (P greater than 0.10) between ULO and sham-operated ram lambs during the period immediately following ULO. 3. Chronically, testosterone concentrations were not significantly different (P greater than 0.10) between ULO and sham-operated ram lambs. 4. After hCG injection, the testosterone response of chronic ULO ram lambs was approx. half the response of the sham-operated ram lambs. 5. These data indicate that a rapid and sustained compensatory response of basal secretion of testosterone but not hCG-inducible testosterone secretion occurred in the ULO'd ram lambs.     

Relationship between in vitro sperm functional tests and in vivo fertility of rams following cervical artificial insemination of ewes with frozen-thawed semen

Several procedures have been proposed to assess structural and functional characteristics of cryopreserved ram semen but none so far have yielded consistent relationships with in vivo fertility. The objectives of this study were to evaluate several sperm function tests as potential markers of in vivo ram fertility (determined by pregnancy rate in ewes) using frozen-thawed semen. In experiment 1, frozen-thawed straws (n=3 per ram) of semen from three high and three low fertility rams were assessed using fluorescent microscopy for (1) progressive motility, (2) viability and, (3) acrosomal status. In experiment 2, frozen-thawed straws (n=3 per ram) of semen from 18 rams of known fertility were analysed using either computer-assisted sperm analysis (CASA) for eight motion characteristics or flow cytometric staining for: (1) viability and acrosomal status, (2) plasma membrane status and capacitation-like changes, and (3) live cells following an osmotic resistance test (ORT). In experiment 3, platelet-activating factor (PAF) was isolated from straws (n=2 per ram) of semen using high-pressure liquid chromatography (HPLC) and quantified using HPLC-tandem mass spectrometry for 18 rams. In experiment 1, no association was found between motility, viability (% live) or acrosomal status (% damaged, % intact and % reacted) and in vivo fertility. In experiment 2, no correlation was found between motility (CASA), viability (% live), acrosomal status (% live, % live intact and % reacted), capacitation status (% capacitated, % non-capacitated), plasma membrane stability (% dead) and % live cells following ORT and ram in vivo fertility. In experiment 3, there was no relationship between PAF content in spermatozoa and ram fertility. In conclusion, we were unable to relate the in vivo fertility of rams with in vitro functional tests of their frozen-thawed semen and suggest that the fertility of a given semen sample cannot easily be quantified using available in vitro tests.     

An automated method for the evaluation of ram libido in real mating conditions

We investigated if sexual behaviour of rams can be assessed with an electronic Alpha-Detector (AD) which automatically records mounts of mating rams. To evaluate the rams’ libido (i.e. all sexual activities), we used six intact and six vasectomised rams in pen tests in three different seasons (late spring, autumn and early spring). The pen tests consisted of 30-min visual observations of each ram placed in a group of six Merino ewes (three ewes in oestrus and three ewes not in oestrus). In the pen tests, sexual behaviour was recorded and divided into two categories: pre-copulatory and copulatory. For validation purposes, during the pen tests the 12 rams were equipped with the AD and the number of times the 18 oestrous ewes were mounted were counted over a period of 3 days. Of the 1191 mounts visually identified in the six 30-min sessions, 1026 were recorded automatically by the AD (i.e. 94%). The paddock test is an automated method consisting of the same rams wearing an AD and placed in a flock of ~250 Merino ewes on two occasions (late spring (spring 1) and early spring of the following year (spring 2)), their copulatory activities were automatically recorded over a 5-day period. The results of the pen tests in the three seasons revealed no difference between the two types of rams (breeding v. detecting rams). Based on live observations high correlations (r=+0.81, P<0.003 for breeding and r=+0.76, P<0.02 for detecting rams) were found between pre-copulatory and copulatory behaviours. The libido of the two types of rams measured in pen tests showed high repeatability across the three seasons (83 and 75%, P<0.05 for copulatory and pre-copulatory behaviours, respectively). When measured automatically in paddock tests over two consecutive springs, even higher repeatability was observed in both breeding (94%; P<0.01) and detecting rams (97%; P<0.004) in the number of mounts. In addition, high correlations (+0.89<r<+0.94) between copulatory behaviours, as measured by live observations, and those measured by the AD were obtained. The automatic measurement of ram libido in paddock tests appears to be more reliable than pen tests and far less time consuming. We therefore recommend this automated method to estimate the libido of rams. In addition, this method can be used at any season of the year provided that ewes in oestrus are present in the flock.     

Melanin concentrating hormone (MCH) in the cerebrospinal fluid of ewes during spontaneous oestrous cycles and ram effect induced follicular phases

The melanin-concentrating hormone (MCH) is a neuropeptide synthesized by neurons of the lateral hypothalamus and incerto-hypothalamic area that project throughout the central nervous system. The aims of the present report were: (1) to determine if MCH levels in cerebrospinal fluid (CSF) of ewes vary between the mid-luteal and the oestrous phase of spontaneous oestrous cycles; and (2) to study if MCH levels in CSF of ewes vary acutely during the follicular phase induced with the ram effect in anoestrous ewes. In the first experiment, CSF was collected from 8 adult ewes during spontaneous oestrous and during the mid-luteal phase (8-10 days after natural oestrus). In the second experiment, performed during the mid non-breeding season, a follicular phase was induced with the ram effect. After isolating a group of 16 ewes from rams, CSF was obtained from 5 of such ewes (control group). Three rams were joined with the ewes, and samples were obtained 12h (n=6) and 24h (n=5) later. In Experiment 1, there were no differences in MCH concentrations in CSF measured during the mid-luteal phase and spontaneous oestrus (0.14 ± 0.04 vs. 0.16 ± 0.05 ng/mL respectively). In Experiment 2, MCH concentrations tended to increase 12h after rams introduction (0.15 ± 0.08 vs. 0.35 ± 0.21 ng/mL, P=0.08), and increased significantly 24h after rams introduction (0.37 ± 0.15 ng/mL, P=0.02). We concluded that MCH concentration measured in the CSF from ewes increased markedly during the response to the ram effect but not during the natural oestrous cycle of ewes.     

Response of anestrous ewes to the ram effect after follicular wave synchronization with a single dose of estradiol-17beta

Anestrous ewes respond to the introduction of rams with either an ovulation within 2-3 days that may be followed by luteal phases of normal or short length, with delayed ovulations (5-6 days later), or with the luteinization of follicles. The aim of this work was to study the relationship between the growth status of the largest follicle present when rams are introduced and the type of ovarian response in non-treated ewes and in ewes treated with estradiol-17beta before ram introduction. Thirteen anestrous Corriedale ewes were divided into 2 groups: E2 (n = 7) and C (n = 6). The E2 ewes received a single dose of 50 microg estradiol-17beta 5 days before the introduction of the rams to synchronize the onset of their follicle waves, while C ewes remained untreated. When the rams were introduced, all E2 ewes had the largest follicle in a growing stage in contrast with the C ewes (3 out of 6; P < 0.05). Five C and 4 E2 ewes ovulated after the introduction of the rams (Day 3.4 +/- 0.4 for C vs. 4.8 +/- 0.3 for E2 ewes, respectively, P < 0.05). Only one ewe from each group developed a normal luteal phase: 4 C and 3 E2 ewes had short luteal phases. One C ewe and 2 E2 ewes had short luteal phases originating from follicles that did not ovulate. After the first luteal phase, all ewes returned to anesirus without a second ovulation or luteal phase. The remaining E2 ewe did not ovulate or show any changes in progesterone serum concentrations. We conclude that the growth status of the largest follicle alone does not determine the ovarian responding pattern of anestrous ewes to the ram effect.     

Seasonal variation in testes size in Polish Merino rams and its relationship to reproductive performance in spring

Observations relating testes size to body weight in growing and adult Polish Merino rams were performed during 3 consecutive years in a flock of 1700 pedigree ewes. October-born ram lambs were measured for testis diameter (TD), scrotal circumference (SC), and body weight (BW) at 100 (January), 150 (March), 180 (April), 240 (June), 365 (October), and 480 d (February) of age. In adult rams (n = 63 to 73 per year) BW and SC were measured in March, May (prior to breeding), August and October. Semen quality was evaluated before each breeding season, and the results of controlled reproductive performance (2 natural services to designated rams) were analyzed. Indices were applied to relate testis size to body weight: TD BW , SC BW and TW BW . Heritability estimates of testes indices in ram lambs at 100 to 240 d and at 480 d of age ranged from 0.21 to 0.36 (SE = 0.067 to 0.090). In adult rams SC measurements within months in consecutive years were repeatable (r = 0.55 to 0.68; P相似文献   

Opposite variations of two epididymal components and blood plasma testosterone in two breeds of rams.

1. Testicular volume (T Vol), blood plasma testosterone (T) concentration, seminal plasma alpha-glucosidase (alpha-G) specific activity, L-carnitine (L-C) concentration as well as semen characteristics were compared in eight Finnish Landrace (F) and eight Suffolk (S) rams throughout 21 months. 2. Only T Vol and T exhibited a typical seasonal variation in both breeds, whereas L-C, alpha-G and live sperm output presented a seasonal profile only in S rams. 3. L-C and alpha-G variations were opposite to those of T in S rams, while they fluctuated in F rams throughout the entire experiment, as did live sperm output. 4. Only the number of ejaculates and T were significantly higher in F rams (3.50 +/- 0.08 in 5 min and 7.62 +/- 0.40 ng/ml) than in S rams (2.30 +/- 0.05 in 5 min and 5.5 +/- 0.30 ng/ml); these two characteristics might therefore be considered as two indexes of sexual activity in rams. 5. By contrast, among all characteristics measured, only alpha-G was significantly higher in S rams than in F rams (1.33 +/- 0.04 vs 0.77 +/- 0.03 mU/mg proteins); this result, as well as seasonal alpha-G profile present in only S rams, allowed us to conclude that alpha-G might be considered as an additional index of seasonal reproduction in rams.     

Effect of melatonin implants on secretion of luteinizing hormone in intact and castrated rams

Rams were treated with melatonin implants in 2 experiments designed to examine the control of reproductive seasonality. In Exp. 1, rams (n = 12) were allocated to 3 treatment groups: 2 groups were treated with 2 melatonin implants per ram for 4 months from 11 November (N) and 9 December (D) and the remaining group was untreated (C). The seasonal increase in luteinizing hormone (LH) pulse frequency and testes size was advanced in Groups N and D. A second seasonal cycle in LH secretion and testes size occurred in Groups N and D after melatonin implants became exhausted. In Exp. 2, rams (n = 20) were allocated to 4 treatment groups: 10 rams were castrated on 6 October and 1 group of entire rams (EM) and one group of castrated rams (CM) were treated with 2 melatonin implants per ram each month from 3 November until 8 January. The other group of entire rams (EC) and castrated rams (CC) was untreated. An increase in LH pulse frequency occurred after castration. Melatonin treatment increased LH pulse frequency in entire rams and reduced LH pulse frequency in castrated rams. The results demonstrated that the advanced reproductive development as a result of treatment with melatonin implants was due to an effect of melatonin on the hypothalamic pulse generator to increase LH pulse frequency. The ability of melatonin to influence LH pulse frequency in entire and castrated rams indicated that an effect of melatonin on the hypothalamic pulse generator is independent of testicular steroids.     

Linear dose-response relationship of erythrocyte enzyme-activity mutations in offspring of ethylnitrosourea-treated mice

The specific activity of 10 erythrocyte enzymes was measured to detect gene mutations in F1 offspring of male mice treated with 3 different doses of ethylnitrosourea (ENU). After administration of ENU or of the solvent (controls), the (101/El X C3H/El)F1 hybrid males were mated to untreated T-stock females. No enzyme-activity mutant was found in 3610 F1 offspring of the control group. After treatment of postspermatogonial germ-cell stages, 1 mutant in 1125 F1 offspring of males treated with 160 mg ENU/kg body weight, and 2 mutants in 1319 F1 offspring of a 250-mg/kg group were observed. After treatment of spermatogonia, 9 enzyme-activity mutants in 4247 F1 offspring of males treated with 80 mg ENU/kg body weight, 15 mutants in 3396 F1 offspring of a 160-mg/kg group, and 9 mutants in 1402 F1 offspring of a 250-mg/kg group were detected. The mutation frequencies in spermatogonia were significantly different from that of the controls (P less than 0.01). The dose-response curve was found to be linear. The frequencies of enzyme-activity mutations are comparable to those of recessive specific-locus mutations determined in the same experiments. Enzyme-activity mutants with reduced activity as well as mutants with enhanced activity were found. Genetic and biochemical characterization of enzyme-activity mutants was routinely performed. In inter se crossings of heterozygotes, no offspring expressing a third phenotype other than the wild type and the heterozygote were found in approximately half of the mutation studies. The recovered mouse mutants might be used as animal models to study corresponding genetic diseases in humans.     

Effect of breed and age on sexual behaviour of rams

The objective of this study was to highlight the problems that arise during the reproduction between thin-tailed rams and fat-tailed ewes. At the same time, particular emphasis laid on the influence of sheep breed, sheep age, time after ram introduction and day of the ewe estrus cycle on ram and ewe sexual behaviour. Rams were subjected to sexual performance tests by being individually exposed to 12 ewes for 3 h daily, 19 consecutive days. The 16 rams of the experiment were separated according to their age (9 and 21 months old) and breed (Chios and Karagouniki), and the 96 ewes of Chios fat-tailed breed, were divided by age (9 and 21 months old). The main characteristics of courtship behaviour, like sniffing, nudging, flehmen response and following were recorded and studied in detail. Mature Chios rams, which were the only one with previous experience of Chios ewes, exhibited higher rates of sexual interest per ewe than the other rams (P < 0.05). On the other hand, rams sniffed and nudged more young than mature ewes (P < 0.05), probably due to the fact that young ewes did not express intense symptoms of estrus. Young rams exhibited substandard sexual interest towards mature ewes, when they first came in contact with them (P < 0.05). In general, Karagouniki thin-tailed rams exhibited reduced rates of mating behaviour when they courted with Chios fat-tailed ewes in comparison with Chios rams (P < 0.05). Moreover, as the time after ram introduction passed, the frequency and duration of sexual behaviour components decreased (P < 0.001). Finally, the effect of the day of the experiment was only significant in the case of sniffing, which increased during the first 2 days and then declined and stabilized (P < 0.01). As it was demonstrated, ram age and ram breed played a fundamental role in the exhibition of sexual interest elements.     

Short term, repeated exposure to rams during the transition into the breeding season improves the synchrony of mating in the breeding season

The ram effect is widely used in Mediterranean breeds of sheep but its use in temperate genotypes is restricted by breed seasonality. However, ewes from these highly seasonal genotypes are sensitive to stimulation by rams close to the onset of the natural breeding season. In this study we developed a pre-mating protocol of repeated, short-term exposure to rams (fence-line contact or vasectomised rams) beginning during late anoestrus and continuing into the breeding season. We hypothesised that this pre-mating protocol would synchronise the distribution of mating of North of England Mule ewes during the breeding season above that observed in ewes isolated from rams prior to mating. Ram-exposed ewes were given contact with rams (Experiment 1: fence-line; FR, n = 94 and Experiment 2: vasectomised rams; VR; n = 103) for 24 h on Days 0 (10 September), 17 and 34 of the experiment. Control ewes (Experiment 1; FC, n = 98 and Experiment 2; VC; n = 106) remained isolated from rams prior to mating. In Experiment 2, a subset of VR (n = 35) and VC ewes (n = 35) were blood sampled twice weekly to monitor their pre-mating progesterone profiles. At mating, harnessed entire rams were introduced, 17 or 16 days after the last ram exposure (Experiments 1 and 2) and raddle marks were recorded daily. The median time from ram introduction to mating was reduced in ewes given both fence-line and vasectomised ram contact (P < 0.001), leading to a more compact distribution of mating and lambing (At least P < 0.01). In the blood sampled VR ewes, there was a progressive decline in the number of days from ram exposure to the onset of dioestrus (at least P < 0.05). This observation indicates that the cycles in VR ewes became increasingly synchronised over the pre-mating period, a pattern not evident in VC ewes. In conclusion, repeated, short-term exposure of ewes to rams during the transition into the breeding season is an effective method of synchronising the distribution of mating during the breeding season.