Reversibility of germinative and endocrine testicular function after long-term contraception with a GnRH-agonist implant in the tom—a follow-up study

A significantly reduced gonadotropin and testosterone secretion is a well-described result of long-term administration of GnRH agonists in the male dog and cat. To date, no data are available about the duration of efficacy and the reversibility of treatment-induced effects after long-term treatment with a 4.7 mg deslorelin implant. Seven healthy male European Shorthair cats (3.2 ± 0.5 kg, 1–6 years) were treated with a 4.7 mg deslorelin implant. Blood samples (testosterone, T), testicular volume, penile spines, and mating behavior were recorded once weekly. Considering T > 0.5 ng/mL as the biological endpoint, mean duration of efficacy was 78.8 ± 12.9 weeks (range: 61.7–100.7 weeks) with T concentrations increasing rapidly after the last T less than 0.1 ng/mL (basal) (P < 0.0001), and pretreatment T concentrations being reached after 3 weeks. Testicular volume rapidly increased after the first increase of T (P < 0.001) with pretreatment testicular volume being reached after 6.9 ± 3.4 weeks (5–11 weeks). “Normal” libido reoccurred 88.7 ± 12.4 weeks after treatment, and “normal” mating behavior was observed even later. Fertile matings occurred 7 to 42 weeks after the last T less than 0.1 ng/mL with a mean of 4.0 ± 0.0 kittens, and 13.6 to 47.6 weeks afterwards testicular histology revealed normal spermatogenesis. The present data confirm that the use of slow-release GnRH-agonist implants containing deslorelin in tomcats represents an effective and safe reversible alternative for long-term contraception; however, as number of animals is low, further fertility trials are recommended.     

Effects of long-term treatment with the GnrH agonist deslorelin (Suprelorin®) on sexual function in boars

Immunization against GnRH has been proven effective for boar taint removal, and long-term treatment with GnRH analogues has been shown to suppress GnRH dependent reproductive processes in several species. This study was conducted to treat boars (n = 5) with Suprelorin®, i.e., an implant that contains 4.7 mg of the long-acting GnRH analogue deslorelin, and to test the effects on sexual function. Insertion of the implant occurred at the age of 5 weeks and animals were observed until market age at 26-27 weeks. Surgically castrated (n = 4) and intact boars (n = 3) served as controls. Testes growth was markedly reduced and steroidogenesis (testosterone, estrone, estrone sulphate, estradiol 17β) as well as spermatogenesis suppressed in 4 of 5 GnRH treated boars, respectively. The remaining fifth boar resumed testes growth after week 17 of age and had high hormone concentrations when tested at weeks 26 and 27. Restoration of spermatogenesis was observed at 34 weeks of age. There were no effects of treatment on general health, nor were there local inflammatory reactions. Results indicate that suppression of sexual functions in boars due to long-term treatment with the GnRH agonist deslorelin through an implant such as Suprelorin® is possible and can last for several months up to market age; thus it has potential as an alternative to other methods used for boar taint removal. Because the maximum duration of suppression seems to vary between boars, further studies are necessary to refine the treatment.     

Reversible suppression of sexual activity in tomcats with deslorelin implant

The aim of the study was to assess the efficacy of using a Gn-RH agonist implant (deslorelin, 4.7 mg, Suprelorin) to control sexual activity of male cats and reestablishment of sexual function after the implant removal 4 mo after placement. Using a control group (Group 1, n = 5), 22 domestic tomcats were given the implant subcutaneously in the region of the right shoulder blade and were then divided into two treatment groups. Animals in Group 2 (n = 14) were observed from the date of implant surgery and the observation lasted for 4 mo. In Group 3 (n = 8) all animals were monitored from the date of implant surgery. Then, after 4 mo, all implants were removed and the toms were observed for a further 4 mo. In all animals during their first visit and then in 1-mo intervals, changes in testosterone concentrations were assessed before (T0) and 4 h after (T4) human chorionic gonadotropin (HCG) administration and testis size was measured. In all tomcats, semen collection was performed, using an electroejaculator, in the course of the first visit and then in 2-mo intervals or at the end of observation. Total sperm count was determined in each semen sample. Two to four animals were castrated at weeks 4, 8, 12, 16, 20, 24, 28 and 32 and histologic assessment of the testes was performed.By evaluation of 200 cross sections of seminiferous tubules, the degree of spermatogenic suppression was assessed and animals in Groups 2 and 3 were assigned into groups according to most tubules with the most developed germ cell observed: G1, spermatocytes; G2, round spermatids; G3, elongating spermatids and G4, elongated spermatids. The mean area of Leydig-cell nuclei was calculated.In animals in Group 2, suppression after implant insertion was monitored. T4 concentrations, testis size, and total sperm count gradually decreased (P < 0.01; P < 0.01; and P < 0.05, respectively) within 4 mo after implantation. Histologic evaluation showed a high individual variation in the degree of suppression of spermatogenesis.In animals in Group 3, the implant was removed 4 mo after insertion and the return of sexual activity was monitored. Within 4 mo, T4 concentration and total sperm count increased to the physiological values of intact toms. Testes gradually increased in size and within 4 mo of implant removal almost reached pretreatment size. According to histologic evaluation of the seminiferous tubules, as early as 1 mo after implant removal, all animals were assigned to G4, with most tubules containing elongated spermatids as the most developed germ cells.Treatment with the long-term subcutaneous Gn-RH agonist implant was well tolerated and no adverse treatment-related effects were noted.These results demonstrated efficacy of 4.7 mg deslorelin implant (Suprelorin) with high variability of the effect onset in tomcats. Furthermore, the study revealed a strong need for complex examination, including testis size measurement, monitoring of hormonal changes, spermatological analysis and histologic evaluation, to declare the animal infertile. After the implant removal, all observed parameters confirmed the reversibility of the method and gradual return of sexual activity in toms.     

Reproductive seasonality and the effect of the GnRH agonist deslorelin as a contraceptive in captive male Black Flying-foxes (Pteropus alecto)

Effective contraception would enhance genetic management of captive Pteropus species, which typically breed well in captivity. Male reproductive seasonality was monitored (15-mo interval) in captive P. alecto (6 controls and 5 treated with 4.7 mg deslorelin). In untreated males, there were seasonal changes in testicular volume, body weight and testosterone secretion; testicular volume and body weight peaked in February and March, respectively, whereas testosterone concentration remained >5 ng/ml before rising (P < 0.001) to 24.9 ± 3.6 ng/ml (mean ± SEM) in April. However, there was no corresponding change in sperm quality, and seminal vesicle gland (SVG) secretions remained present in ejaculates. In treated males, testosterone concentration had an initial ‘flare’ response (mean ± SEM peak: 19.95 ± 3.27 ng/ml) before declining (P < 0.001) by 32 d to basal levels, where it remained. In these males, there was reduced sperm motility after 1 mo (P < 0.001) and the absence of SVG secretions after 4 mo. However, aspermic ejaculates were first recorded 5 mo post-treatment. At 10 mo after treatment, spermatogenesis was still disrupted, when membrane-intact, but non-motile sperm were present in two individuals. Motile sperm were first recovered from one of these males 13 mo after deslorelin treatment. We concluded that captive P. alecto males: (a) had seasonal reproductive changes in testicular volume, body weight and testosterone secretion; (b) produced motile, membrane-intact sperm and SVG secretions throughout the year; and (c) had a rapid decline in testosterone concentration and consequent suppression of testicular function for at least 5 mo following deslorelin administration.     

Reversible downregulation of endocrine and germinative testicular function (hormonal castration) in the dog with the GnRH-Agonist Azagly-Nafarelin as a removable implant “Gonazon”; a preclinical trial

Downregulation of anterior pituitary GnRH-receptors by application of a slow release GnRH-implant offers an effective and reversible alternative to surgical castration of the male dog.Aim of the present study was to test the efficacy and the underlying mechanisms of a new non-biodegradable controlled-release device implant (Gonazon^®, Intervet, containing 18.5 mg of the GnRH-agonist Azagly-Nafarelin). Eight male beagle dogs were implanted s.c. at the para-umbilical region. In four dogs implant removal was after 180 days (group 1), in the other four dogs after 365 days (group 2). Eleven weeks after implantation availability of LH was reduced (p < 0.0001) by 70%. After an initial increase lasting for about 4 days, testosterone (T) and estradiol (E2) concentrations decreased (p < 0.0001) to basal levels within 17.5 ± 8.4 days. Size of testes was decreased by about 82% after 17 weeks, size of prostate by about 46% after 5 weeks (p < 0.0001). Five to 7 weeks after implantation all dogs were aspermic.Testosterone and estradiol concentrations, together with testicular and prostatic size remained suppressed in all dogs in group 1 and one dog of group 2 until implant removal. The other three dogs of group 2 escaped from down-regulation between 223 and 324 days.Effects on the availability of LH, T, E2 and on testicular and prostatic size were fully reversible after implant removal or escape from down-regulation. In six dogs semen quality was back to pre-treatment values after about 29 weeks, however, one dog developed oligozoospermia while another one stayed azoospermic, probably due to an obstruction within the epididymal duct.     

Early pregnancy diagnosis of captive Amur tigers (Panthera tigris altaica)

From January 2015 to February 2017, urine samples was collected from twelve captive female Amur tigers during both their mating (or receptive) periods and the first four weeks after mating at the Hengdaohezi Feline Breeding Center (FBC) in China. A radioimmunoassay was applied to determine the progesterone levels in the urine samples. The relationship between progesterone levels and pregnancy in Amur tigers was assessed according to whether the tigers gave birth in the end. Results showed that average progesterone levels increased significantly (P < .05) after mating for all female tigers having parturition. And the minimum value of progesterone levels of them during the fourth week after mating was consistently higher than 0.21 ng/mL, the maximum value in mating period of all female tigers. However, there were no significant differences in progesterone levels between mating period and the first four weeks after mating of the female tigers having no parturition. Therefore, early pregnancy in captive Amur tigers can be diagnosed when urine progesterone concentrations attain a level higher than 0.21 ng/mL for seven days in a row after mating.     

Induction of fertile oestrus in the bitch using Deslorelin, a GnRH agonist

Oestrus induction in various canine breeds was attempted in 32 bitches. A group of 8 bitches were treated 80–160 d following their previous oestrus (G1) whereas a second group of 24 bitches (G2) were implanted 200–590 d following their previous oestrus. The treatment for each bitch consisted in one Deslorelin implant (Suprelorin®4,7mg, Virbac, France), inserted subcutaneously in the post-umbilical region. Ovulation, pregnancy rate and litter size were recorded. All bitches came in heat 4.3 ± 1.4 d after implantation (2–7 d). Ovulation was reported in 62.5% in G1 and 87.5% in G2. One bitch refused mating and since no AI was performed, she was not considered for further analysis. Pregnancy was obtained in 25% in G1 versus 78.3% in G2. Mean litter size was 6.7 ± 3.5 puppies (1–14). Luteal failure was suspected in 3 bitches, two that remained non-pregnant and one which aborted 58 d post-ovulation since the owner refused progesterone supplementation. Deslorelin implants can therefore be considered as a valuable alternative to induce fertile oestrus in bitches in anoestrus. Follow-up of the luteal phase is recommended, since some bitches might encounter luteal failure.     

乙醛脱氢酶基因ALDH2对雄性小鼠生育能力的影响

本研究使用乙醛脱氢酶2(ALDH2)基因全敲除的雄性C57BL/6(B6)小鼠（Musmusculus domesticus）,通过分析不同周龄小鼠的睾丸脏器系数、睾丸组织细胞形态、精子运动参数、配种后母鼠产仔数及子代雄雌比等生育指标,探讨ALDH2基因敲除对雄性小鼠生育能力的影响。结果表明,与野生型（WT）小鼠相比,5、7、10周龄ALDH2基因敲除型（KO）雄性小鼠睾丸脏器系数显著降低（P <0.05）;睾丸组织细胞间质变大,精子活率显著降低（P <0.05）;产仔数和雄雌比显著降低（P <0.05）。本研究为揭示乙醛脱氢酶ALDH2基因在雄性小鼠生殖中的作用提供了一定的基础。     

Ovarian activity reversibility after the use of deslorelin acetate as a short-term contraceptive in domestic queens

The objective was to evaluate ovarian activity reversibility in domestic queens after short-term contraceptive treatment with deslorelin acetate. Ten mature queens were used. In all queens, the estrous cycle was evaluated every 72 h by vaginal cytology (VC) and behavior assessments. When queens had VC characteristic of interestrus or diestrus, one deslorelin acetate implant (4.7 mg) was placed in the subcutaneous tissue of the interscapular region (day of insertion = Day 0). Thereafter, VC was performed every 48 h and on Day 90, implants were removed. At Day 100, estrus and ovulation were induced with 100 IU eCG (im), followed by 100 IU hCG (im), 84 h later (Day 103.5). Queens were ovariohysterectomized on Day 106. Corpora lutea (CL) were counted, oviducts were flushed, and oocytes were identified, isolated and stained to assess viability. In all queens, blood samples for plasma progesterone concentrations were collected once a week, from Days −21 to 106. After deslorelin acetate application, four queens had VC and behavior typical of estrus, and one ovulated. Furthermore, ovulation occurred in three queens that did not have VC or behavior consistent with estrus. After the initial ovarian stimulation, all females had anestrous VC during the deslorelin treatment period. Implants were readily removed. Following implant removal, all females responded to treatments to induce estrus and ovulation. There were (mean ± SEM) 13.1 ± 5.5 CL and 8.1 ± 5.5 oocytes per queen; the oocyte recovery rate was 56.8 ± 25.4% and all recovered oocytes were viable. We concluded that deslorelin acetate can be used as a reversible short-term contraceptive in domestic cats, because estrus and ovulation were successfully induced following implant removal.     

Effects of the GnRH analogue deslorelin implants on reproduction in female domestic cats

The aim of the present study was to investigate the safety and efficacy of deslorelin, a GnRH agonist, implants in suppressing estrus behavior and matings in a controlled ambient environment in feline queens in the presence of a tomcat. Local and utero-ovarian side effects of deslorelin implants were also investigated. The queens were housed in groups and assigned to one of three treatments: group 1 received 9.5 mg deslorelin implants (N = 14), group 2 received 5 mg megestrol acetate tablets and 9.5 mg deslorelin implants (N = 7), and group 3 were given placebo implants (N = 7). All implants were placed subcutaneously cranial to the interscapular region under xylazine hydrochloride sedation. Ovarian activity was monitored by fecal estradiol (E₂) analyses. The animals were observed daily and checked individually at three-day intervals for behavioral signs of estrus. After 18.5 mo of trial, queens were ovariohysterectomized, and ovaries and uteri were weighed and evaluated histologically. E₂ levels were significantly lower in group 1 and 2 than in group 3 with an average of 128.48 ± 19.97 ng/g, 90.44 ± 7.16 ng/g and 283.26 ± 39.21 ng/g, respectively, excepting the first week of treatment. After inserting implants an initial estrus-like increase in fecal E₂ concentrations occurred in all treated queens except one female in group 2. Ovarian and uterine weights were significantly different among the groups (P < 0.01), and were lowest in groups 1 and 2. Primordial and primary follicle numbers were significantly higher in groups 1 and 2 than in group 3 (P < 0.001). Endometrial gland, antral follicle, and corpus luteum (CL) numbers were highest in group 3 (P < 0.01, 0.001, and 0.001, respectively) compared with groups 1 and 2. Deslorelin implants successfully suppressed estrus behavior and E₂ secretion in queens for 18.5 mo of the study period. Further investigations are needed to demonstrate the effects of GnRH agonists on ovarian interstitial tissue.     

In vitro effects of estradiol-17β, monobutyl phthalate and mono-(2-ethylhexyl) phthalate on the secretion of testosterone and insulin-like peptide 3 by interstitial cells of scrotal and retained testes in dogs

The objective was to determine the effects of estradiol-17β, monobutyl phthalate (MBP) and mono-(2-ethylhexyl) phthalate (MEHP) on testosterone and insulin-like peptide 3 (INSL3) secretions in cultured testicular interstitial cells isolated (enzymatic dispersion) from scrotal and retained testes of small-breed dogs. Suspension cultures were treated with estradiol-17β (0, 10, and 100 ng/mL), MBP (0, 0.8, and 8 mmol/L) or MEHP (0, 0.2, and 0.8 mmol/L) for 18 h, in the presence or absence of 0.1 IU/mL hCG. Testosterone (both basal and hCG-induced) and INSL3 (basal) concentrations were measured in spent medium. Effects of estradiol-17β, MBP, and MEHP on testosterone and INSL3 secretions were not affected (P > 0.15) by cell source (scrotal versus retained testis); therefore, data were combined and analyzed, and outcomes reported as percentage relative to the control. In testicular interstitial cells, basal testosterone secretion was increased (P < 0.01) by 100 ng/mL estradiol-17β (130.2 ± 10.6% of control). Among phthalates, 0.2 and 0.8 mmol/L MEHP stimulated (P < 0.01) basal testosterone secretion (135.5 ± 8.3% and 154.6 ± 12.9%, respectively). However, hCG-induced testosterone secretion was inhibited (P < 0.01) by 8 mmol/L MBP (67.7 ± 6.0%), and tended to be inhibited (P = 0.056) by 0.8 mmol/L MEHP (84.5 ± 5.6%). Basal INSL3 secretion was inhibited (P < 0.01) by 8 mmol/L MBP (73.6 ± 6.8%) and 0.8 mmol/L MEHP (76.9 ± 11.3%). In conclusion, we inferred that estradiol-17β and certain phthalate monoesters had direct effects on secretions of testosterone and INSL3 in canine testicular interstitial cells, with no significant difference between scrotal and retained testes.     

The use of a deslorelin implant (GnRH agonist) during the late embryonic period to reduce pregnancy loss

Embryonic and fetal mortality reduce reproductive performance of lactating dairy cows. The objectives of this study were to reduce pregnancy loss by administering a deslorelin implant (GnRH agonist) during the late embryonic period, to reduce follicular growth, induce accessory corpora lutea, and increase plasma progesterone concentrations. Lactating dairy cows received an implant containing 2.1 mg of deslorelin (Deslorelin group; n = 89) or no treatment (Control group; n = 92) on Day 27 of pregnancy. Pregnancy, ovarian structures and plasma progesterone concentrations were determined on Days 27 and 45, and pregnancy was re-confirmed on Day 90. On Day 45, mean +/- S.E.M. numbers of class 2 (6-9 mm; 0.72+/-0.19) and class 3 (> or = 10 mm; 0.86 +/- 0.12) follicles for cows in the Deslorelin group were lower (P < 0.01) than the numbers of class 2 (1.90 +/- 0.18) and class 3 (1.92 +/- 0.12) follicles for cows in the Control group. On Day 45, the number of accessory corpora lutea for cows in the Deslorelin group (1.80 +/- 0.07) were greater (P < 0.01) than for cows in the Control group (1.31 +/- 0.07). On Day 45, plasma progesterone concentration was increased (P < 0.01) for cows in the Deslorelin group (8.03 +/- 0.33 ng/mL) compared to cows in the Control group (6.40 +/- 0.31 ng/mL). Pregnancy losses did not differ between Days 27 and 45 and Days 45 and 90 for cows in the Control (15.2 and 11.0%, respectively) and Deslorelin groups (20.2 and 10.5%, respectively). However, in the Deslorelin group, pregnancy loss between Days 45 and 90 was lower (P < 0.05) for cows that formed an accessory CL (0%) compared to cows that did not form an accessory CL (16.1%).     

Testosterone undecanoate in the functional compartments of the male reproductive tract

Background

Assessment of testosterone undecanoate’s (TU) presence in the functional compartments of the male reproductive tract has never been performed despite the evidence that its documented beneficial effect in male infertility might be mediated through an epididymal action and this study was set to examine this possibility.

Materials and methods

In 18 normozoospermic volunteers TU has been administered (40 mg t.i.d.) for 6 days with serum measurements of TU, total testosterone (T), DHT, E2, SHBG, FSH, LH, and PRL before and at the end of medication. Steroid hormones (T, E2, and TU) were also assayed in seminal plasma. In a selected group of 7 men with previously diagnosed non-obstructive azoospermia TU, T, and E2 were assayed in the extracts of testicular biopsy material taken before ICSI and at the end of the same medication.

Results

A marked rise of serum DHT (average 148%, P < 0.001) has been found after treatment, whereas T, E2, FSH, LH, SHBG, and PRL did not significantly change. Measurable amounts of TU were found in the serum of all men but only in 6 cases in seminal plasma (11.1 ± 8.0 ng/mL) and all of them in semen delivered 7–8 h after the last TU capsule was taken. In dilution fluid from testicular tissue extracts, no detectable amounts of TU were found whereas mean values of 92.5 ± 54.3 pg/mL and 43.8 ± 16.3 ng/mL for E2 and T were observed. Positive correlations among TU and E2, T or DHT concentrations were found in serum samples (P < 0.01, 0.02, and 0.002) as well as between E2 and T (P < 0.01), E2 and DHT (P < 0.001), or T and DHT (P < 0.001).

Conclusion

It is concluded that TU was identified and measured for the first time in seminal plasma of a fair percentage (33%) of men on this medication and was associated in all men with a marked rise of DHT concentration, a known epididymal function promoter, in the absence of an effect on pituitary and gonadal activity. On this evidence, it appears that a beneficial effect of TU on epididymal function may be a distinct possibility.     

Morphofunctional alterations in testicular cells of deslorelin-treated boars: an immunohistochemical study

In this study we thoroughly scrutinized testes morphology and investigated whether treatment of recipient boars with gonadotropin-releasing hormone (GnRH)-agonist deslorelin could alter the expression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD), luteinizing hormone receptors (LHRs), and androgen receptors (ARs) in testicular cells. An implant containing 4.7 mg of the GnRH-agonist deslorelin was subcutaneously inserted into crossbred male pigs at 91 and 147 days of age. Testicular traits, morphology of the testes, the proteins' expression, and testosterone concentration in blood plasma were analyzed in all boars after slaughter at 175 days of age. Histological analysis revealed significant alterations in both the interstitial tissue and seminiferous tubules of experimental animals after 28 and 84 days of deslorelin treatment. The intensity of the AR immunostaining within the testis appeared as a function of the severity of testicular dysgenesis. Time-dependent action of deslorelin on the expression of LHR and 3beta-HSD in Leydig cells was also detected. Staining for LHR and 3beta-HSD was very weak or the Leydig cells were immunonegative. Concomitantly, a significant decrease in plasma testosterone level was found in both groups of deslorelin-treated boars when compared with the control group. This is the first report showing the cellular distribution of AR, LHR, and 3beta-HSD in testicular cells of deslorelin-treated boars. It is concluded that morphological and immunohistochemical studies are important for the evaluation of testicular histoarchitecture and steroidogenic function. Subsequently, the endocrine control of reproduction in the GnRH-agonist deslorelin-treated males will be better understood.     

Concentrations of altrenogest in plasma of mares and foals and in allantoic and amniotic fluid at parturition

Treatment with the progestin altrenogest is widely used in pregnant mares. The fact that foals born from healthy mares treated with altrenogest until term suffered from neonatal problems raises the question of direct effects of altrenogest on vital functions in the neonate. We have therefore investigated altrenogest concentrations in maternal and neonatal blood plasma and in fetal fluids. Pregnant mares were treated with altrenogest orally once daily (0,088 mg/kg bodyweight, n = 7) or left untreated (n = 8) from 280 d of gestation until foaling. Altrenogest concentration was determined in plasma of the mares, their foals and in amniotic and allantoic fluid. The concentration of altrenogest in plasma from treated mares (2.6 ± 1.0 ng/mL) was significantly lower than in plasma from their foals immediately after birth (5.6 ± 1.9 ng/mL; p < 0.05), but was significantly higher than in their fetal fluids (amniotic fluid: 0.4 ± 0.1 ng/mL; p < 0.05; allantoic fluid: 3.0 ± 1.5 ng/mL). Altrenogest was undetectable in maternal and fetal plasma and fetal fluids of control pregnancies at all times. Altrenogest concentration in plasma of foals from treated mares was strongly correlated to the altrenogest concentration in plasma of their dams (r = 0.938, p < 0.001) and in amniotic (r = 0.886, p < 0.001) and allantoic fluid (r = 0.562, p < 0.05). A significant decrease in altrenogest concentration between the time periods 0-15 min, 30-120 min, and 180-360 min after parturition was seen in the plasma from foals born to altrenogest-treated mares. In conclusion, our data demonstrate that altrenogest reaches the equine fetus at high concentrations.     

Effect of xylazine,isoxsuprine, and lidocaine on Doppler sonographic uterine and umbilical blood flow measurements in cows during the last month of pregnancy

The maternal portion of the bovine placenta receives blood mainly from the uterine arteries (AUT) and the fetal portion from the umbilical arteries (AUM). Placental perfusion is crucial for fetal development and undergoes adaptive changes during pregnancy according to the fetal requirements. One goal of this study was to investigate changes in Doppler sonographic measurements of blood vessels that supply blood to the placenta in cows during the last 4 weeks of pregnancy. Another goal was to examine how these measurements are affected by three drugs commonly used in cows at the time of parturition. Nine cows underwent Doppler sonographic examination of the AUT ipsilateral and contralateral to the pregnant horn and one AUM three times per week during the last 4 weeks of gestation. This was followed by the randomized administration of one of the three following experimental drugs per day: isoxsuprine (200 mg/cow, iv), xylazine (2 mg/100 kg, iv), and lidocaine for epidural anesthesia (100 mg/cow). Doppler sonographic examination was repeated 30 minutes after medication. Maternal pulse rate increased during the study period (P < 0.001), and the diameter of the contralateral AUT was smaller in the last week before birth than in the two preceding weeks. The resistance index (RI) of the ipsilateral AUT was smaller in the last week than in the first 2 weeks of the study period. Uterine blood flow volume increased after isoxsuprine by 5% and after epidural anesthesia by 6% (both P ≤ 0.05) and decreased after xylazine by 10% (P < 0.001). Isoxsuprine was the only drug that elevated the blood flow volume in the AUM (P ≤ 0.05). Xylazine increased the RI of both AUT (both P < 0.001) and significantly reduced maternal and fetal pulse frequencies, whereas isoxsuprine significantly reduced the RI of both AUT and the AUM and increased maternal and fetal pulse frequencies. The results reported that Doppler sonographic measurements of uterine and AUM change little in the last month of pregnancy in the cow. Isoxsuprine and epidural anesthesia with lidocaine have the potential to improve uterine perfusion.     

Weight Loss and Leptin Changes in Individuals with Type 2 Diabetes

WILLIAMS, KATHERINE V., MONICA MULLEN, WE1 LANG, ROBERT V. CONSIDINE, AND RENA R. WING. Weight loss and leptin changes in individuals with type 2 diabetes. Obes Res. Objective To identify variables associated with leptin change in subjects with type 2 diabetes after 3 weeks and 20 weeks of weight loss. Research Methods and Procedures Subjects with type 2 diabetes treated with diet or sulfonylureas (n = 54) were enrolled in a 20-week behavioral weight control program. Sulfonylureas were stopped ≥2 weeks before study entry. Seven subjects who restarted sulfonylureas after week 3 had their data analyzed separately after this point. Results Leptin, fasting plasma glucose, and insulin levels were measured at baseline and at 3, 10, and 20 weeks. After 3 weeks, subjects lost 2.7±2.0 kg (p<0.001), and had significant decreases in leptin (5.2±7.0 ng/mL, p<0.001), fasting plasma glucose (1.8±1.8 mmol/L, p<0.001), and insulin (23±60 pmol/L, p<0.03). Between week 3 and week 20, subjects lost an additional 6.3±4.4 kg (P<0.001), but had no further changes in leptin. The primary determinants of leptin change at all time-points were weight loss and initial leptin level. Changes in insulin were not related to changes in leptin after controlling for the effects of weight loss. At week 20, more recent weight loss (week 10 to week 20) was as strong a predictor of overall change in leptin as overall weight loss (baseline to 20 week). Subjects who restarted sulfonylureas had an increase in both leptin levels (+1.9±9.0 ng/mL, p<0.05) and insulin levels (+23±65 pmol/L, p<0.05), despite significant overall weight loss (-7.4±4.0 kg, p<0.01). Initial changes in leptin (0 weeks to 3 weeks) did not affect subsequent ability to lose weight. Discussion Both short- and long-term changes in weight had an effect on leptin changes in individuals with type 2 diabetes. Although physiological insulin changes did not independently influence changes in leptin concentration with weight loss, increases in insulin levels with sulfonyl-urea therapy were associated with increases in leptin levels despite weight loss.     

Testicular testosterone: Estradiol ratio in domestic cats and its relationship to spermatogenesis and epididymal sperm morphology

The phenomenon of teratozoospermia in felids is not fully understood. In this study, we investigated the testicular androgen:estrogen balance in domestic cats and correlated these data with epididymal sperm morphology and the degree of spermatogenic activity. During spring and summer, testes and blood samples were obtained from 37 mixed-breed domestic cats (12 to 48 mo). The epididymal sperm were harvested and evaluated for sperm counts, motility, and morphology. Distal cytoplasmic droplets were not considered a defect, and samples were considered normozoospermic if they contained more than 60% normal sperm (N = 25) or teratozoospermic if they contained less than 45% normal sperm (N = 12). The testicular and serum concentrations of testosterone (T) and 17β-estradiol (E2) were determined with an enzyme immunoassay. The gonadosomatic index and epididymal sperm numbers and motility did not differ between groups. The percentage of normal sperm was higher in normozoospermic (74.3 ± 2.0, mean ± SEM) than in teratozoospermic samples (43.1 ± 1.4). The most prevalent sperm defects in the teratozoospermic group were abnormal acrosomes (9.7 ± 2.0) and bent midpieces (12.2 ± 2.0) or tails (24.0 ± 2.7) with cytoplasmic droplets. Histomorphometric data were similar between groups, although there was a lower Leydig cell nuclear volume in teratozoospermic samples. Normozoospermic samples contained a higher percentage of haploid cells and had a higher index of total spermatogenic transformation than teratozoospermic samples. Serum concentrations of T (0.5 ± 0.1 vs. 0.8 ± 0.4 ng/mL) and E2 (9.5 ± 1.2 vs. 11.4 ± 2.3 pg/mL) and testicular T concentrations (471.6 ± 65.3 vs. 313.4 ± 57.6 ng/g) were similar between groups. However, compared with normozoospermic samples, teratozoospermic samples had higher testicular E2 concentrations (8.5 ± 3.6 vs. 5.4 ± 0.5 ng/g) and a lower T:E2 ratio (31.8 ± 4.1 vs. 87.2 ± 11.6). There were significant correlations between testicular E2 values and percentages of normal sperm (r = −0.55) as well as those with primary sperm defects (r = 0.58) or abnormal acrosomes (r = 0.64). The T:E2 ratio was also correlated with meiotic index (r = 0.45) and percentage of normal sperm (r = 0.58). In conclusion, a high testicular E2 concentration and a reduced T:E2 ratio were significantly associated with higher ratios of abnormal sperm types, suggesting that the balance between androgens and estrogens is an important endocrine component in the genesis of teratozoospermia in felids.     

Endocrine effects of the GnRH antagonist, acyline, in domestic dogs

Gonadotrophin-releasing hormone (GnRH) antagonists may have a future role in the control of canine reproductive function. In this study, the effects of a single dose of the potent GnRH antagonist, acyline, on serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) were evaluated in male dogs. Blood samples were drawn before (Day −1) and after (30, 60, and 90 min, 3, 6, 9, 12, and 24 h, and 3, 6, 9, 14, 22, and 29 d) treatment with acyline (330 μg/kg, sc); serum concentrations of FSH, LH, and T varied throughout the study period (P < 0.01, <0.05, and <0.01, respectively). Gonadotrophins decreased below pretreatment concentrations 60 min after injection, whereas T took 90 min to decrease below baseline (P > 0.05). Follicle-stimulating hormone, LH and T decreased until Day 9, when they reached their nadir at 2.0 ±1.1 ng/mL (P < 0.01), 1.2 ± 0.2 ng/mL (P > 0.05), and 0.5 ± 0.2 ng/mL (P < 0.05), respectively. Both gonadotrophins and T began increasing on Day 14 after treatment, although FSH and T serum concentrations still remained below baseline on that day (P > 0.05). Follicle-stimulating hormone and T rebounded above baseline on Day 29, whereas LH reached concentrations were similar to baseline at this time (P > 0.05). No local or systemic side effects were detected in any dog following acyline treatment. In conclusion, a single acyline treatment safely and reversibly decreased serum gonadotrophin and T concentrations in dogs for 9 d.