The use of a 4.7 mg deslorelin slow release implant in male dogs in the field.

OBJECTIVE: Slow-release GnRH agonist implants (SRI) are used for reversible medical downregulation of testicular function in male dogs as an alternative to surgery. The 4.7 mg deslorelin SRI should reduce testosterone after 6-8 weeks and induce castration-like effects for 6 months (mon). However, some individual variation is described in the field in regard to onset and duration of effect. For this reason, we aimed to study the effects of the 4.7 mg deslorelin SRI in a larger cohort. MATERIAL AND METHODS: In total 50 intact, healthy male dogs (12-48 months, mon; 9-40 kg) were treated with a 4.7 mg deslorelin SRI into the umbilical area (TG, n=45) or served as untreated controls (CG, n=5). CG dogs were surgically castrated after measurement of testicular dimensions and blood sampling for testosterone. In TG, SRIs remained for 5 mon in place and subsequently 3-7 male dogs were surgically castrated at removal (week, W 0) or 1, 2, 3, 4, 5, 6, 7, 8 or 10 weeks later. Examination parameters were testicular dimensions (before treatment, at 4, 8, 12 W, 5 mon, weekly until castration), testosterone (before treatment, at 8 W, 5 mon, castration) and testicular histology (castration). RESULTS: Whereas examination parameters did not differ between CG and TG before treatment, testicular volume and testosterone was significantly reduced at all time points during treatment. In all but 3 (8 W) and 2 male dogs (5 mon) testosterone was basal during treatment before removal, whereas the parameters were significantly reduced compared to pre-treatment in the respective dogs. After implant removal, testosterone and testicular volumes increased. However, different to earlier studies, the "restart" was more variable with individual basal testosterone until W7, but also physiological testosterone concentrations in W2. Similarly, histological testicular findings at castration were quite variable: besides an arrest on spermatogonia and spermatocytes, elongated spermatids with normal spermatogenesis were found in individual dogs. CONCLUSION: Our study confirms the efficacy of the deslorelin SRI, but also individual variation especially regarding reversibility of effects on endocrine and germinative testicular function. CLINICAL RELEVANCE: Deslorelin SRIs offer a suitable alternative to surgical castration with individual variation to be considered when used in clinical practice.

The Efficacy of a 3ß-Hydroxysteroid Dehydrogenase Inhibitor for the Termination of Mid-Term Pregnancies in Dogs.

Progesterone (P4) is the only hormone needed to maintain pregnancy in dogs. Therefore, a competitive inhibitor of 3ß-hydroxysteroid dehydrogenase (3ß-HSD) could be a safe and effective option to terminate pregnancy by inhibiting P4 synthesis. To address this hypothesis, we investigated the efficacy of trilostane (TRL), a competitive inhibitor of 3ß-HSD, in terminating pregnancy in dogs. Twenty-one dogs between days 30 and 38 of pregnancy were randomly assigned to one of two treatment groups (trilostane (TRL) and aglepristone (AGL)) and an untreated control (CON) group (n = 7 dogs each). Fetal heart rates (FHRs) (measured at 12 h intervals) and serum P4 concentrations (measured at 6 h intervals) were evaluated. The pregnancy termination rates were 0% and 100% in the TRL and AGL groups, respectively. The decrease in the FHR in the TRL and AGL groups was significantly lower than that observed in the CON group. There was a marked decrease in P4 concentrations in the TRL group 6, 54, and 102 h after the initiation of treatment. The luteal expression of StAR appeared to be weaker in the AGL group than the CON group. In conclusion, although a treatment-induced decrease was observed in plasma P4 concentrations, a seven-day TRL treatment alone was not effective in terminating pregnancies. Further studies are needed on the effects of the prolonged administration of TRL with varying doses and frequencies for the termination of mid-term pregnancy in dogs.

Plasma concentration and uterine and ovarian expressions of insulin-like growth factor-2 in dogs with cystic endometrial hyperplasia-pyometra.

The objective of this study was to investigate the plasma concentrations of insulin-like growth factor-2 (IGF-2) as well as its expression in the uterus and ovary of healthy dogs and those with cystic endometrial hyperplasia (CEH)-pyometra complex. Group 1 (n = 10) included bitches with open cervix pyometra, while Group 2 (n = 7) consisted of clinically healthy bitches in dioestrus. The number of IGF-2 immunopositive interstitial cells was significantly higher in Group 1, whereas in Group 2 there were only two cases in which a few cells were IGF-2 immunopositive. IGF-2 immunopositivity was observed in the endometrial glandular epithelium in both groups. Additionally, interstitial fibroblasts and macrophages in the endometrium were also positive in Group 1. The concentration of plasma IGF-2 was higher in Group 1 than in Group 2 (P < 0.05). The concentration was positively correlated with IGF-2 expression in the endometrial glands (r = 0.926; P < 0.001) in Group 1. However, a negative correlation was present between plasma IGF-2 concentration and IGF-2 expression in the interstitial endocrine cells of the ovary in Group 1 (r = -0.652; P < 0.05). The results suggest that IGF-2 plays an important role during the inflammatory process occurring in bitches with CEH-pyometra complex as well as in the endometrium of healthy bitches in dioestrus.

Gene expression profiling of the canine placenta during normal and antigestagen-induced luteolysis.

The domestic dog is the only domestic animal species that does not produce steroids in the placenta and instead relies on luteal steroids throughout pregnancy. Nevertheless, the canine placenta is highly responsive to steroids, and withdrawal of progesterone (P4) affects the feto-maternal unit, initializing the parturition cascade. Similar effects can be observed during antigestagen-induced abortion. Here, aiming to provide new insights into mechanisms involved in the termination of canine pregnancy, next generation sequencing (NGS, RNA-seq) was applied. Placental transcriptomes derived from natural prepartum and antigestagen-induced abortions were analyzed and compared with fully developed mid-gestation placentas. The contrast "prepartum luteolysis over mid-gestation" revealed 1973 differentially expressed genes (DEG). Terms associated with apoptosis, impairment of vascular function and activation of signaling of several cytokines (e.g., IL-8, IL-3, TGF-ß) were overrepresented at natural luteolysis. When compared with mid-term, antigestagen treatment revealed 135 highly regulated DEG that were involved in the induced luteolysis and showed similar associations with functional terms and expression patterns as during natural luteolysis. The contrast "antigestagen-induced luteolysis over prepartum luteolysis" revealed that, although similar changes occur in both conditions, they are more pronounced during natural prepartum. Among P4-regulated DEG were those related to immune system and cortisol metabolism. It appears that, besides inducing placental PGF2α output, both natural and induced P4 withdrawal is associated with disruption of the feto-maternal interface, leading to impaired vascular functions, apoptosis and controlled modulation of the immune response. The time-related maturation of the feto-maternal interface needs to be considered because it may be clinically relevant.

Effect of oral misoprostol, alone or in combination with aglepristone, on mid-term pregnancy termination in cats.

OBJECTIVES: This study aimed to determine the efficacy and safety of oral misoprostol (MIS) administration in the induction of mid-term pregnancy termination in cats. METHODS: Twenty-eight cats that were pregnant for 30-40 days were allocated to four groups. The aglepristone (AGL) group (n = 7) received 10 mg/kg SC aglepristone q24h for two consecutive days. In the AGL+MIS group (n = 7), AGL (as administered in the AGL group) and MIS (200 µg/cat PO q12h until the start of abortion) were administered. The MIS200 (n = 7) and MIS400 groups (n = 7) received MIS (200 or 400 µg/cat misoprostol, respectively) alone PO q12h until the start of abortion. Blood samples were collected at the start of treatment (d0), 4 days after the start of treatment (d4) and on the day of complete abortion/end of administration (dA/d7). RESULTS: The efficacy of the treatment was 71.4% in the AGL group, 100% in the AGL+MIS group, 0% in MIS200 group and 57.4% in MIS400 group (P = 0.004). No significance was found in relation to the interval from treatment to the start/end of abortion and the duration of abortion in all groups. The most observed side effect was vomiting in both groups administered MIS, particularly in the MIS400 group (56.7%). Progesterone (P4) concentrations were reduced during the abortion, but not to basal levels, in all groups. P4 concentrations were significantly lower at dA/d7 in the MIS400 group compared with the AGL and AGL+MIS groups (P = 0.002). CONCLUSIONS AND RELEVANCE: The results obtained from this study showed that low doses of MIS do not induce abortions in cats but increase the effect of AGL. Although higher doses could terminate pregnancies, this also causes intense unwanted side effects. Therefore, the use of MIS alone as an abortifacient in cats is not recommended. For mid-term pregnancy termination in cats, the combination of misoprostol and aglepristone provides a more effective abortifacient than using either of them alone.

Effect of deslorelin on testicular function, serum dihydrotestosterone and oestradiol concentrations during and after suppression of sexual activity in tom cats.

Objectives The aim of the study was to evaluate the efficacy of a 4.7 mg deslorelin implant in tom cats. Methods Nine mature male cats were included in the deslorelin group and five cats in the control group. Before the study started, all cats were confirmed to have distinct sexually dimorphic behaviour. Blood samples were taken on the implantation day, at day 7 and at day 15, then monthly, in order to measure serum dihydrotestosterone (DHT) and 17beta(ß)-oestradiol concentrations. The deslorelin group (n = 9) was divided into two subgroups: five cats (cats 1-5) were neutered in the postimplantation period during suppression of sexually dimorphic behaviour, and four cats (cats 6-9) were neutered after re-expression of sexually dimorphic behaviour. The control group cats (n = 5) were castrated without administration of the implant. Results Sexually dimorphic behaviours ceased within a mean ± SD of 13-58 days (23.30 ± 14.17) after implantation. DHT concentration decreased within 30 days. The mean duration of suppression was 26.5 ± 7.42 months and reactivation coincided with increased DHT values reaching preimplantation concentrations within 1 month. 17ß-oestradiol concentrations significantly correlated with DHT concentrations ( P <0.01). For cats castrated during suppression of sexual behaviour, the length of the long axes of the nuclei of Leydig cells, the diameter of seminiferous tubules and the height of the epithelium of the seminiferous tubules did not change until 3-6 months after implantation, whereas at 12 and 32 months the measured values were even lower than in the control group. For cats castrasted after reactivation, the length of long axes of the nuclei of Leydig cells and the diameter of seminiferous tubules approached the values of the control group between 4 and 6 months after reactivation. Conclusions and relevance A deslorelin implant (4.7 mg) suppresses sexually dimorphic behaviour in tom cats without any side effects and with full reversibility; however, duration of suppression is highly individual.

Tarantula cubensis extract alters the degree of apoptosis and mitosis in canine mammary adenocarcinomas.

In the present study, 13 clinical cases of canine mammary adenocarcinoma were evaluated in order to understand the effect of Tarantula cubensis extract (TCE) on tumor tissue. Punch biopsies were taken from the tumors before treatment with TCE. Subcutaneous injections of TCE were administered three times at weekly intervals (3 mL per dog). Between days 7 and 10 after the third injection, the tumor masses were extirpated by complete unilateral mastectomy. Pre- and post-treatment tumor tissues were immunohistochemically assessed. The expression of B-cell lymphoma 2 (Bcl-2) was found to be higher in pre-treatment compared to post-treatment tissues (p < 0.01) whereas Ki-67 expression was lower in post-treatment tissues (p < 0.01). No significant differences in fibroblast growth factor or vascular endothelial growth factor expression were observed between pre- and post-treatment tissues (p > 0.05). The apoptotic index was determined to be low before treatment and increased during treatment. These results suggest that TCE may be effective for controlling the local growth of canine mammary adenocarcinoma by regulating apoptosis.

Right intercostal insertion of a Veress needle for laparoscopy in dogs.

OBJECTIVE: To evaluate right intercostal Veress needle (VN) insertion for laparoscopy in dogs. STUDY DESIGN: Longitudinal cohort study. ANIMALS: Female dogs (n = 56). METHODS: The VN was inserted in the last palpable right intercostal space, either 1/3 (Group T; 28 dogs) or mid distance (Group H; 28 dogs) from the xiphoid cartilage to the most caudal extent of the costal arch. Problems encountered during VN insertion and injuries were recorded, graded, and compared between groups, and also between the first and last 20 insertions. RESULTS: Pneumoperitoneum was successfully achieved by VN insertion in 49 (88%) dogs after a single (45 dogs) or 2nd attempt (4 dogs). Frequency of complications was as follows: 20 grade 1 (subcutaneous emphysema, omentum, or falciform injuries); 6 grade 2 (liver or spleen injury), and 1 grade 3 complication (pneumothorax occurred). No significant difference was found between the 2 groups or between the first and last 20 dogs. CONCLUSIONS: Right intercostal VN insertion facilitates pneumoperitoneum in dogs with few consequential complications. No significant difference was found between entry sites; however, the mid distance insertion site in the last palpable intercostal space with dog positioned in dorsal recumbency is likely to result in less complications.

Laparoscopic ovariectomy in dogs: comparison between single portal and two-portal access.

OBJECTIVE: To compare surgical times and perioperative complication rates of single portal access and 2-portal laparoscopic ovariectomy (LapOVE) in dogs using a bipolar vessel sealer/divider device, and to evaluate the performance of novice laparoscopists for right ovariectomy. STUDY DESIGN: Controlled clinical trial. ANIMALS: Female dogs (n=42). METHODS: Dogs were divided into groups: 1=single portal and 2=2 portal. LapOVE was performed using a 5 mm vessel sealer/divider device and a 10 mm operating laparoscope (Group 1) or a 5 mm laparoscope (Group 2). Dog characteristics (weight, body condition score, ovarian ligament fat score), operative time, and perioperative complication rate were compared between groups. Right ovariectomy duration was evaluated for 2 novice laparoscopists. RESULTS: No significant difference was found in mean total surgical time between group 1 (21.07 min/s) and group 2 (19.06 min/s). Factors significantly affecting times included body condition scores, ovarian ligament fat score, ovarian bleeding, and surgeon expertise. Minor complications (bleeding from ovaries or after splenic trauma) occurred and were similar in both groups. Bleeding was correlated to body condition score and ovarian ligament fat score. Interindividual differences were found among surgeons for right ovariectomy time. CONCLUSIONS: Single portal access LapOVE using vessel sealer/divider device is feasible, safe, and does not significantly increase total surgical time in comparison with 2-portal approach. Laparoscopic skills may play a role in ability to perform single portal LapOVE. CLINICAL RELEVANCE: LapOVE can be performed using single portal access.

Induction of abortion in queens by administration of cabergoline (Galastop) solely or in combination with the PGF2alpha analogue Alfaprostol (Gabbrostim).

The efficacy of cabergoline solely or combined with a PGF2alpha analogue in inducing abortion at different stages of pregnancy was investigated in 18 queens. The queens were assigned to two treatments: Group I ( n=10 )-cabergoline (15 microg/kg; daily, orally) and Group II ( n=8 )-cabergoline (15 microg/kg; daily, orally) combined with alfaprostol (10 microg/kg; every other day, subcutaneously). Each group was divided into two subgroups according to the duration of pregnancy when treatments started: Group IA ( n=8 ) included queens from Days 34 to 42 after mating. Group IB cats ( n=2 ) started treatments on Day 45 post-mating. Similarly, the combination of cabergoline and PGF2alpha analogue was first given to Group IIA ( n=6 ) from Days 25 to 40 of pregnancy and to Group IIB ( n=2 ) on Days 45 and 47, respectively. Termination of pregnancies was successful in all cats of Group IA, while treatments failed in both cats of Group IB, even though seven and eight treatments, respectively, were given. Mean (+/-S.D.) plasma progesterone concentrations before the start of treatments were 85.0+/-12.3 nmol/l and decreased within 3 days to 8 nmol/l and subsequently to basal values, when the queens aborted (Group IIA, n=6 ) or gave birth prematurely (Group IIB, n=2 ). When abortions failed (Group IB, n=2 ), progesterone concentrations remained elevated (16.9 and 9.8 nmol/l). Duration of combined therapy during late pregnancy in Group IIB ( n=2 ) lasted about 10 days. In both cases, premature birth occurred and the kittens died within 16 h after birth. Overall, treatments starting on Days 25-42 of pregnancy (Groups IA and IIA) had abortion rates of 100%. The average duration of treatments was 5.6+/-1.5 days (range, 3-8). Side effects seen were vomiting and occurred in 6 of the 109 (5.5%) treatments. In conclusion, pregnancies were successfully terminated in the second trimester of feline pregnancy by daily application of cabergoline solely or combined with the PGF2alpha analogue alfaprostol (given every other day). Cabergoline alone was ineffective in inducing abortion at later stages of pregnancy.