Comparative quantitative assessment of GnRH- and LH-receptor mRNA expression in the urinary tract of sexually intact and spayed female dogs.

Ovariectomy interrupts the regulatory loop in the hypothalamus-pituitary-gonad axis, leading to a several-fold increase in gonadotropin levels. This rise in hormonal secretion may play a causal role in ovariectomy-related urinary incontinence. The purpose of this study was to examine the effect of ovariectomy in bitches on the expression of GnRH- and LH-receptors in the lower urinary tract, and assess the relationship between receptor expression and plasma gonadotropin concentrations. Plasma gonadotropins were measured in 37 client-owned bitches. Biopsies were harvested from the mid-ventral bladder wall in all dogs, and from nine further locations within the lower urinary tract in 17 of the 37 animals. Messenger RNA of the LH and GnRH receptors was quantified using RT-PCR with the TaqMan Universal PCR Master Mix. Gonadotropins were measured with a canine-specific FSH-immunoradiometric assay and LH-radioimmunoassay. The hierarchical mixed ANOVA model using MINITAB, Mann-Whitney U-test, unpaired means comparison and linear regressions using StatView were applied for statistical analyses. Messenger RNA for both receptors was detected in all biopsy samples. Age was negatively correlated to mRNA expression of the LH and the GnRH receptors. A relationship between the mRNA values and the plasma gonadotropin concentrations was not established. Evaluation of results within each of the biopsy locations revealed greater LH-receptor expression in the proximal second quarter of the urethra in spayed bitches than in intact bitches (P=0.0481). Increased mRNA expression of LH receptors in this location could possibly play a role in the decrease in closing pressure of the urethra following ovariectomy.

Review of contraception in ungulate species.

Most ungulate species are herd animals. In captivity, and increasingly so in the wild, space constraints limit natural behaviors associated with group dynamics, possibly resulting in inbreeding and/or overpopulation. This situation has necessitated research regarding contraception of various species of hoofstock. Differing management situations mandate different contraception protocols to achieve optimal results. Fertility control in hoofstock has been achieved through a number of different contraceptive methods predominantly surgical sterilization, mechanical contraception, synthetic steroid hormones, and immunocontraception. In this study successes and limitations of these techniques are reviewed. Zoo Biol 26:311-326, 2007. (c) 2007 Wiley-Liss, Inc.

The effect of GnRH analogs on urinary incontinence after ablation of the ovaries in dogs.

After removal of the ovaries approximately 20% of dogs develop urinary incontinence. Removal of the gonads results in estrogen deficiency and chronic elevation in the production and secretion of FSH and LH. The gonadotrophins may directly or indirectly, adversely affect the sphincter function of the urethra. Estrogen replacement therapy and treatment with sympathomimetics, such as ephedrine or phenylpropanolamine (PPA), are effective only in some of the affected dogs, and many of these subsequently become nonresponsive. Since the role of the elevated gonadotrophins has not been elucidated, we used depot preparations of GnRH analogues to down-regulate gonadotrophins once or twice in 13 ovariectomized (ovx), incontinent dogs, which were either refractory to alpha-adrenergics (n=11) or in which alpha-adrenergics were contraindicated (n=2). Dogs were treated with leuprolide, deslorelin, buserelin or triptorelin. In 7 dogs treatments with GnRH analogues alone (n=11) resulted in continence for 50-738 days (mean 247). In all dogs except one, where GnRH treatments did not resolve the incontinence completely, additional treatment with phenylpropanolamine was successful. With additional treatment of phenylpropanolamine complete continence was restored for 21-367 days (mean 159). All treatments caused long-term reduction of circulating FSH and LH concentrations to very low or undetectable levels. No adverse effects of treatments were observed.

Induction of contraception in some African wild carnivores by downregulation of LH and FSH secretion using the GnRH analogue deslorelin.

The GnRH analogue deslorelin, in long-acting biocompatible implants, was used as a contraceptive in 31 cheetahs (13 females and 18 males), 21 African wild dogs (15 females and 6 males), 10 lionesses and four leopards (three females and one male). A dose of 12 or 15 mg deslorelin was administered to lions, whereas 6 mg deslorelin was administered to the other species. Monitoring consisted of observations, measurement of plasma progesterone and testosterone concentrations, vaginal cytology and evaluation of semen and sex organs. Deslorelin induced contraception in lionesses for 12-18 months, and in female cheetahs and leopards for a minimum of 12 months after treatment. Two male cheetahs had no viable spermatozoa or detectable plasma testosterone 21 months after treatment with deslorelin. Female wild dogs responded less consistently and one bitch conceived 4 weeks after implantation. However, in nine bitches, mating could be postponed until the next breeding season. Male dogs responded consistently and the contraception was effective for approximately 12 months. Although lionesses and cheetahs may become attractive to males for a few days after treatment, mating was not observed. No side-effects or behavioural changes were noted, indicating that deslorelin is a safe drug to use for the contraception of the species described. Males remain fertile for the first 6 weeks after the insertion of implants and should be separated from cyclic females during this period.

Assessment of cabergoline as a reproductive inhibitor in coyotes (Canis latrans).

The efficacy of three oral formulations (gelatin capsule, tablet, oil base) and five dosages (50, 100, 250, 500, 1000 microg) of cabergoline to disrupt reproduction in coyotes (Canis latrans) was evaluated. The type of formulation used had no effect on plasma progesterone and prolactin concentrations or on mean litter size. No adverse side effects (for example, vomiting, anorexia, diarrhoea) were observed despite the use of doses of up to 20 times the therapeutic dose used for domestic dogs and cats. All coyotes treated with 50, 100, 250 and 500 microg cabergoline whelped, but plasma progesterone concentrations in these coyotes were lower (P < or = 0.07) than in control animals at day 7 after treatment. Ten of 11 females treated with 1000 microg cabergoline whelped, but progesterone concentrations in these coyotes were lower than in control animals up to day 14 after treatment (P < or = 0.04). Dosages of 1000 microg cabergoline decreased blood serum prolactin (P < or = 0.10) and progesterone (P < or = 0.06) concentrations, but apparently failed to decrease progesterone below the threshold necessary to maintain pregnancy in all but one animal. However, progressive inhibition of prolactin and progesterone with increasing doses of cabergoline indicated that higher dosages might be effective in coyotes. Survival of pups born to cabergoline-treated females was not different (P < 0.001) from that of pups born to control females, but mean litter size was smaller for females treated with cabergoline (P < or = 0.073) than for the control females. Although all cabergoline treatments in this study were ineffective at preventing reproduction in coyotes, progressive inhibition of prolactin and progesterone with increasing dosages of cabergoline indicates that higher doses might be effective in preventing reproduction in coyotes. However, the physiological differences from other canine species in dopamine D2 receptors and mechanisms of luteal support may ultimately prevent the use of cabergoline for reproductive control in coyotes.

Suppression of the oestrous responses of bitches to the GnRH analogue deslorelin by progestin.

Studies were undertaken in Australia and Belgium to determine whether the initial pro-oestrous-oestrous responses of anoestrous bitches to treatment with deslorelin administered in a s.c. implant were inhibited by progestin treatment. Thirty-nine bitches of mixed breeding were treated daily with 2 mg megestrol acetate kg-1 body weight for 21 (group 1, n = 5) or 14 days (group 2, n = 10), or with 1 mg megestrol acetate kg-1 body weight for 14 days (group 3, n = 10). A deslorelin (6 mg) implant was placed s.c. on day 14 (group 1) or day 7 (groups 2 and 3) of treatment. Bitches not treated with progestin also received a deslorelin implant (group 4, n = 9) or were untreated controls (group 5, n = 9). Signs of pro-oestrus-oestrus were not observed in bitches in groups 1, 2 and 5, but were observed in bitches in groups 3 (4/10) and 4 (9/9). Four bitches in group 4 were mated, two of which became pregnant. The pregnancies failed at about day 40 of gestation and were associated with low plasma progesterone concentrations. Treatment with progestin inhibited the pro-oestrous-oestrous responses of bitches to deslorelin.

Efficacy of the GnRH analogue deslorelin for suppression of oestrous cycles in cats.

The aim of this study was to develop a method for long-term but reversible inhibition of oestrous cycles in female cats by downregulation of GnRH receptors with deslorelin released from a long-acting implant. In a blind study with mature cats (n = 20), a 6 mg deslorelin implant was administered s.c. to ten cats and a placebo implant was administered to ten cats. Occurrence of oestrus and general health were observed daily, and individual faecal samples were collected at 3 day intervals for 14 months and analysed for oestradiol content. All the placebo-treated queens continued to undergo normal oestrous cycles during the study. Oestrus was accompanied by peaks in oestradiol concentrations of > or = 20 ng g-1 faeces. Treatment with deslorelin initially stimulated oestradiol release, which accompanied treatment-induced ovulations. Thereafter, oestradiol concentrations decreased to 1-10 ng g-1 faeces and remained low for extended periods. Observations of small increases in oestradiol concentrations in one cat led to a second treatment with 6 mg deslorelin in five cats on day 155 after first treatment. Faecal oestradiol concentrations remained < 20 ng g-1 faeces in the five single treatment cats for 8.0, 8.5, 11.0 and 14.0 (two cats) months. Cats receiving two implants had the first oestradiol peak > 20 ng g-1 faeces after treatment at 7.5, 11.0 (two cats), 11.5 and 14.0 months. After 14 months, two cats had returned to normal cyclic activity, two had irregular small oestrogen peaks and six showed no cyclic activity. For months 2-5, 6-10 and 11-14, oestrogen values in treated cats were significantly different from control values (P < 0.001, 0.05 and 0.02, respectively). Differences in oestrogen concentration between control cats and cats that were treated twice were significant (P < 0.001) during months 6-10 only. The general health of treated cats was unchanged throughout the study. These results confirm that deslorelin can effectively suppress ovarian activity in domestic cats, but that the duration of suppression varies among individuals.

Control of reproduction and sex related behaviour in exotic wild carnivores with the GnRH analogue deslorelin: preliminary observations.

The GnRH analogue deslorelin, in long-acting implants, was used in an attempt to temporarily control reproduction or aggression in wild carnivores in southern Africa and the USA. In the southern African study, 6 mg deslorelin was administered to cheetahs (eight females, four males), one female leopard and wild dogs (six females, one male) housed in groups, and 12 mg deslorelin was administered to two lionesses. None of the animals became pregnant after deslorelin administration apart from one wild dog that was mated at the initial treatment-induced oestrus. Two wild dogs and one lioness came into oestrus 12 and 18 months after deslorelin administration, respectively, thus demonstrating that the anti-fertility effects of deslorelin are reversible. Two lionesses and four cheetahs underwent oestrus without allowing mating 2-14 days after treatment. Simultaneous administration of progestins to three bitches and one lioness did not suppress oestrus. Male cheetahs had no spermatozoa on day 82 after treatment and did not impregnate two untreated females. Of three untreated female wild dogs housed with treated males, only the first female to enter oestrus (21 days after deslorelin administration) became pregnant. One month after treatment, plasma testosterone concentrations of male dogs were at basal values. In the USA study, three male sea otters that had been treated with 6 mg deslorelin ceased antagonistic behaviour and blood testosterone concentrations and size of the testes were still sharply reduced 24 months after treatment. Male red (n = 7) and grey (n = 5) wolves received 6 mg deslorelin in December 1998 but no effects on seasonal spermatogenesis and behaviour were observed. In a black-footed cat, sperm production, libido and aggressiveness decreased in response to treatment with 3 mg deslorelin and penile spines were not observed within 3 months after treatment, but were observed again 4-6 months later. Treatment of female red (n = 5) and grey (n = 5) wolves with deslorelin in December 1999 triggered preseason oestrus and mating, which were followed by one abortion and one successful pregnancy. Contraception was achieved in female Fennec foxes (n = 7) and two lionesses, which was observed in the foxes by an absence of increases in faecal progesterone concentrations. In two male bush dogs, administration of 3 mg deslorelin once or twice was insufficient to suppress reproductive function or behaviour.

Effectiveness of cabergoline for termination of pregnancy in silver fox (Vulpes vulpes fulva).

Red foxes (Vulpes vulpes) are a major pest species in Europe and Australia. Traditional methods of control such as hunting or poisoning are no longer sufficient or feasible. As with domestic dogs and cats, prolactin (PRL) in the vixen is an essential luteotropin during the second half of gestation. Hence, PRL inhibitors such as cabergoline have been used to induce abortions. Eighteen mated silver fox vixens (three groups of six foxes each) were treated orally with a placebo of paraffin oil (I), or with 15 microg/kg cabergoline in feed once (11) or twice (III), on day 30 (I and II) or days 30 and 32 (III) post-coitum. Blood samples were taken prior to and after treatments and concentrations of PRL and progesterone (P4) were determined. Normal parturitions were observed in five of six, five of six and two of six vixens in groups I, II and III, respectively. In group III plasma concentrations of PRL and P4 decreased significantly but only temporarily. This drop in hormone concentrations was more pronounced in the vixens that did not carry to term. In conclusion, doses in excess of 15 microg/kg of cabergoline are likely to prevent the development of fetuses to term in pregnant vixens.

Zinc serum level in human immunodeficiency virus-infected patients in relation to immunological status.

In human immunodeficiency virus (HIV) infection, serum level of zinc, an important micronutrient for immune function, is frequently diminished. The aim of this study was to determine the zinc status in relation to immunological parameters and disease stage in 79 HIV-1 seropositive patients. The median serum level of zinc was within normal limits (12.5 micromol/L) but in 23% of patients, zinc deficiency was seen. Decreased serum zinc was associated with a low CD4 cell count, high viral load, and increased neopterin and IgA levels. According to current treatment recommendations, the majority of patients received antiretroviral triple therapy. Zinc levels in treated and untreated patients were comparable. Referring to disease stage (CDC classification, 1993), the mean zinc level was highest in stage C and lowest in stage A. In conclusion, even under antiretroviral triple therapy, zinc deficiency is still of great importance in HIV infection, and zinc substitution in zinc deficient individuals should be taken into account to optimize therapeutical success.

Zinc status in patients with alveolar echinococcosis is related to disease progression.

Zinc is an essential trace element for immune function that plays a role in immune response against parasites. To determine a possible relationship between zinc level and disease status in alveolar echinococcosis (AE), we investigated serum concentrations of zinc, immunoglobulin (Ig)E, IgG, and C-reactive protein (CRP) in 40 AE patients and 20 controls. Patients were classified into three groups: group A: patients after curative surgery, group B: patients with stabilized disease, group C: patients with progressive disease. Patients showed significantly higher levels of IgE and IgG than controls. Amounts of IgE and IgG were related to disease severity, achieving highest levels in group C and lowest in group A. Zinc levels were comparable in patients and controls. However, there was an obvious association between zinc concentration and disease severity. Zinc was far below the normal range in group C (median 9.2 micromol/l) and significantly diminished compared to group B and controls. An inverse pattern was seen for CRP. In conclusion, lowered zinc concentration in progressive cases may be caused by enhanced immune activation but consumption of zinc by the growing parasite may also play a role. Furthermore, decreased zinc levels may contribute to the observed immunosuppression in AE.

[Abnormal behavior and adaptation problems in dogs and cats and their pharmacologic control]. / Fehlverhalten und Anpassungsprobleme bei Hund und Katze und deren pharmakologische Beeinflussbarkeit.

Small animal practitioners are increasingly confronted with patients showing adaptation related problems (ARP) which are expressed as disturbed or abnormal behavior (DAB). As a result, practitioners are asked increasingly to euthanize animals which seemingly cannot be socialized. In healthy dogs and cats, three main causes for DAB can be detected: refusal of obedience because of the drive for dominance; anxiety and frustration; and geriatric DAB. Increasingly, disease conditions not readily diagnosed can cause DAB, especially hypothyroidism. Influencing and contributing factors to DAB are breed, sex, experiences as a puppy, behavior of owners, changes in the pet's environment. ARPs may also cause disturbances in the condition of skin and fur, e.g. atopic dermatitis, pruritus sine materia, lick granuloma, and of the intestinal organs (vomiting, irritated bowel syndrome) and may result in an immune deficiency. Therapeutic approaches include behavioral therapy, surgical or hormonal castration with progestins or antiandrogens, substitution with thyroxin in cases with hypothyroidism, and/or the use of psychopharmaca, most prominently of modern antidepressiva like amitriptyline; buspirone; clomipramine and fluoxetine, but also of selegiline, a mono-aminoxydase inhibitor. These compounds, among other effects, are elevating prolactin levels. This seems to allow to formulate a working hypothesis: in the canine species, prolactin is obviously a hormone enabling socialization; hence all drugs which safely cause an increase in prolactin production might be suitable to manage or control ARPs and DAB in the dog, but also in the cat. Higher levels of prolactin than those required for socialization, as seen in nursing bitches or some clinically overt cases of pseudopregnancy, may cause maternal aggression and can be controlled with prolactin inhibitors, if needed.

Controlled-release products for the control of the estrus cycle in cattle, sheep, goats, deer, pigs, and horses.

This paper describes the estrus cycles of a number of livestock breeds and reviews the controlled-release drug delivery systems that are currently available for the purpose of controlled breeding. The bovine estrus cycle is reviewed in detail, and the estrus cycles of other species are described in a manner that highlights similarities and differences between species. Pertinent formulation and pharmacokinetic information about current drug delivery systems is presented and discussed, and recent advances in this area are also described.

Multicenter clinical comparison of sedative and analgesic effects of medetomidine and xylazine in dogs.

OBJECTIVE: To evaluate analgesic and sedative effects of medetomidine hydrochloride in dogs and to compare effects with those of xylazine hydrochloride. DESIGN: Randomized, controlled trial. ANIMALS: 184 dogs that required sedation or analgesia for completion of minor diagnostic or therapeutic procedures. PROCEDURE: Dogs were sedated with medetomidine, i.v. (750 micrograms/m2 of body surface area) or i.m. (1,000 micrograms/m2) or with xylazine, i.v. (1.1 mg/kg 10.5 mg/lb] of body weight) or i.m. (2.2 mg/kg [1 mg/lb]). Sedative effects were measured by scoring posture and response to noise. Durations of effects were determined by measuring time intervals between drug administration and changes in posture. Analgesic effects were measured by determining toe-pinch pressure needed to elicit a withdrawal response. Clinicians rated sedative and analgesic effects and ease with which diagnostic or therapeutic procedures could be performed. RESULTS: Posture and response to noise scores were significantly higher for dogs given medetomidine, i.m., than for dogs given xylazine, i.m., and for dogs given medetomidine, i.v., than for dogs given xylazine, i.v. Time to regaining sternal recumbency and time to regaining ability to stand were longest after i.m. administration of medetomidine. Toe-pinch pressures were not significantly different among groups. Clinicians rated overall analgesic and sedative effects as excellent significantly more often after administration of medetomidine than after administration of xylazine. Prevalence of adverse effects did not differ among groups. CLINICAL IMPLICATIONS: Medetomidine and xylazine, at doses tested, were effective and safe, but results of subjective measurements indicated that medetomidine provided better sedation and analgesia than did xylazine. Specific alpha 2-adrenergic antagonists (atipamezole, yohimbine) are available for control of adverse cardiovascular effects.

Use of the GnRH analogue, deslorelin acetate, in a slow-release implant to accelerate ovulation in oestrous mares.

In two separate controlled clinical trials, the efficacy and safety of 2.2 mg of the GnRH analogue deslorelin, administered subcutaneously as a short-term implant to normally cycling mares in oestrus with a dominant ovarian follicle more than 30 mm in diameter, were evaluated, using a placebo as a negative control. The oestrous cycle of each mare was followed by teasing, palpation per rectum and transrectal ultrasonography. Follicles were monitored every 24 hours by ultrasonography until ovulation occurred. The mares were either mated naturally or inseminated artificially. In trial 1, 174 mares were treated at six locations in Canada, and in trial 2, 98 mares were treated at three locations in the USA. In trial 1, the treatment with deslorelin reduced the mean (sd) time to ovulation from 84.2 (48.4) hours to 50.2 (19.6) hours (P < 0.001) and in trial 2 it reduced it from 88.8 (40.3) hours to 54.1 (26.5) hours (P < 0.001). In trial 1, the percentage of mares ovulating within 48 hours increased from 37.7 per cent in control mares to 86.1 per cent in treated mares (P < 0.001) and in trial 2 the percentage increased from 26.5 to 80.9 per cent (P < 0.001). In trial 2, the duration of oestrus in the deslorelin-treated mares was reduced from 6.1 days to 4.3 days and the number of matings or artificial inseminations was reduced from 2.5 to 1.7 (P < 0.001). In trial 1, days 12 to 20 pregnancy rates for matings at the treatment oestrus were not different for deslorelin-treated (75.6 per cent) and placebo-treated (66.1 per cent) mares. In trial 2, days 12 to 20 pregnancy rates from matings at the treatment oestrus were lower for deslorelin-treated (58.7 per cent) than for placebo-treated (83.3 per cent) mares (P < 0.05), although pregnancy rates were similar for deslorelin-treated (97.1 per cent) and placebo-treated (95.0 per cent) mares after mating at the second oestrus. In both trials, pregnancy losses due to early or late abortions were within the normally expected range and similar for deslorelin-treated (3.6 and 3.7 per cent, respectively) and placebo-treated (13.4 and 7.5 per cent) mares. The treatments did not cause systemic side effects and local reactions at the implantation sites were slight and of short duration.

Techniques developed for the control of estrus, ovulation and parturition in the East German pig industry: a review.

Centralized administration of the nationalized pig industry in the former German Democratic Republic (East Germany), promoted wide-scale use of assisted reproduction of pigs. The objective was to synchronize reproductive events such as estrus, ovulation, breeding and parturition in a way that would allow for quick penetration of desirable genetic information by timed artificial insemination and the routine use of hygienic measures. Methods developed to control these events were based on extensive basic and applied research, but the resulting pool of unique scientific information was heretofore not published in refereed journals in the English language. We present here a comprehensive review of biotechnological procedures and physiological insights gained by basic and applied research and by their application at the large pig production units.