Review of seven cases of granulosa cell tumour of the equine ovary

This paper reviews the equine granulosa cell tumour (GCT) and describes the clinicopathological features, treatment and outcome in seven cases of GCT in mares. Mares were presented with unilateral ovarian enlargement during the 2007 to 2010 breeding seasons. The mean (sd) age of the mares was 11.7 (5.96) years. Three mares were multiparous barren, three were nulliparous and one was primigravida. Behaviour at presentation was 57 per cent anoestrus, 28 per cent with stallion-like behaviour and 14 per cent with persistent oestrus. All mares had unilateral ovarian enlargement. Six non-pregnant mares had a small and inactive contralateral ovary; the pregnant mare had a single small corpus luteum on the contralateral ovary and was at three-and-a-half months' gestation. Enlarged ovaries measured 7 cm to an estimated 30 cm in diameter. 28 per cent had a multicystic ultrasound appearance, 57 per cent were dense structures and 14 per cent were of mixed appearance. Mean concentrations of progesterone were <1 ng/ml, oestrone sulphate 3.06 (2.32) ng/ml and testosterone 0.58 (0.64) nmol/l in non-pregnant mares. Inhibin was elevated in all non-pregnant cases at 7.6 (12.45) ng/ml. The pregnant mare had concentrations of progesterone 2.5 ng/ml, oestrone sulphate 81.0 ng/ml, testosterone 1.9 nmol/l and inhibin 1.31 ng/ml. Mares demonstrating stallion-like behaviour had a significantly higher (P<0.001) testosterone concentration (1.85 [0.07] nmol/l) than those that did not (0.34 [0.26] nmol/l). Three mares underwent unilateral ovariectomy and resumed cyclic ovarian activity within nine months of surgery.     

Elevated Serum Anti-Müllerian Hormone in an Arabian Mare with Granulosa Cell Tumor

Anti-Müllerian hormone (AMH), produced by granulosa cells, has been reported to be elevated in mares with granulosa cell tumor. A 13-year-old Arabian mare was referred after 3 years of infertility. Rectal and ultrasonographic examination revealed enlargement and multicystic appearance of the left ovary. The mare had substantially higher concentration of AMH (21.6 ng/mL) compared with normal cyclic mares (n = 5; 0.36 ± 0.02 ng/mL). Testosterone (50 pg/mL) concentration was also elevated, but progesterone concentration (0.4 ng/mL) was lowered. Accordingly, the mare was presumptively diagnosed with granulosa-theca cell tumor and a left ovariectomy implemented using ventral midline approach under general anesthesia. Histopathological examination confirmed the diagnosis of granulosa-theca cell tumor. Serum concentrations of AMH decreased to 2 ng/mL and 0.5 ng/mL 2 and 4 weeks after tumor removal, respectively. In conclusion, the current report presents confirmatory evidence that evaluation of serum AMH could be useful for diagnosis of granulosa cell tumor in mare.     

Determination of serum anti‐Müllerian hormone concentrations for the diagnosis of granulosa‐cell tumours in mares

Reasons for performing study: Endocrinological assays are important for evaluation of mares with granulosa‐cell tumours (GCTs), and our research in mares indicates that anti‐Müllerian hormone (AMH) may be a good biomarker for this type of ovarian tumour. Objectives: To evaluate the use of serum AMH concentrations for endocrine diagnosis of GCTs in mares. Methods: Archived serum samples (n = 403) previously assayed for determination of serum inhibin, testosterone and progesterone concentrations (GCT panel) were assayed for serum AMH concentrations using a heterologous enzyme‐linked immunosorbent assay previously validated by our laboratory. For a subset (n = 44) of these samples, a clinical diagnosis of GCT was confirmed by histopathology. Results: Overall, the sensitivity of AMH (98%) for detection of histologically confirmed GCTs was significantly (P<0.05) greater than that of either inhibin (80%) or testosterone (48%) or the combination of inhibin and testosterone (84%). Conclusions: Determination of serum AMH concentrations is a useful biomarker for detection of GCTs in the mare. Potential relevance: Measurement of serum AMH concentrations can be used for diagnosis of GCTs in the mare. As serum AMH concentrations do not vary significantly during the oestrous cycle or pregnancy, interpretation of these results is not confounded by these physiological states.     

Luteoma in two mares treated by ovariectomy

A 15-year-old Paint mare (Case 1) and a 15-year-old Arabian mare (Case 2) were presented for aggressive and undesired behaviour. Reproductive evaluation revealed, initially, an enlarged ovary with a smaller/normal sized contralateral ovary in each case. Granulosa cell tumour (GCT) panel testing revealed elevated inhibin B in both cases and elevated anti-Mullerian hormone (AMH) levels only in Case 1 determined from currently established reference ranges. Based on a presumptive diagnosis of GCT, bilateral standing ovariectomy was elected for both mares. In each case neoplastic tissue consistent with luteoma was detected only on histological examination in the smaller/normal sized left ovary; the right ovary appearing normal. Long-term follow-up was performed 1 year and 5 years post-operatively by telephone. The owners were satisfied with the outcome and each patient demonstrated resolution of aggressive stallion-like behaviour. Each case returned to the owners' intended use. In conclusion, luteoma should be considered as a differential diagnosis for ovarian neoplasms in horses related to behavioural abnormalities, even in normal-sized ovaries.     

Anti‐Muellerian hormone concentration in bitches with histopathologically diagnosed ovarian tumours and cysts

Increased concentrations of Anti‐Muellerian hormone (AMH) can indicate a granulosa cell tumour as shown in women, mares and cows. To investigate AMH to differentiate canine granulosa cell tumour from other ovarian pathologies, we evaluated the ovaries of 63 bitches. Blood serum samples were collected before surgery for AMH analysis. Ovaries were submitted for histopathological examination. Fourteen bitches showed normal ovaries. These bitches had AMH values between 0.12 and 0.99 ng/ml. In 20 bitches ovarian cysts i.e., follicular cysts (n = 8), corpora lutea cysts (n = 7), subsurface cysts (n = 5) were diagnosed. These dogs had AMH values of 0.11–2.09 ng/ml. Bitches with small luteinized follicular cysts had slightly higher AMH values than those without ovarian alteration. In 29 cases ovarian neoplasms i.e., granulosa cell tumour (n = 9), epithelial tumours (n = 16), dysgerminomas (n = 3) and one sarcoma were identified. Anti‐Muellerian hormone values of bitches with an ovarian neoplasm except granulosa cell tumour ranged from 0.18 to 1.18 ng/ml. The AMH values of bitches with granulosa cell tumour ranged from 1.12 to ≤23 ng/ml and were significantly higher (p < .05) than in all of the other bitches. The cut‐off of 0.99 ng/ml gave a sensitivity of 100% and a specificity of 94.44% to diagnose granulosa cell tumour. In conclusion, markedly elevated AMH concentrations in bitches are indicative for a granulosa cell tumour. However, negative testing does not rule out the existence of small one. Differentiation of GCT from luteinized follicular cysts may especially be difficult.     

Granulosa cell tumours in the mare: A review of 52 cases

Granulosa cell tumour (GCT) is reported to be the most common ovarian neoplasm in the mare. A multicentre retrospective study evaluating the medical records of horses undergoing ovariectomy for treatment of a GCT was undertaken, documenting the presenting clinicopathological features, treatment and outcome. Follow‐up information was obtained by telephone questionnaire. Fifty‐two mares were included. Most presented with behavioural/reproductive abnormalities, including stallion‐like behaviour (50%), aggression (31%), prolonged oestrus or nymphomania (19%), and persistent anoestrus (8%). Most (93%) affected ovaries had a multilocular appearance when examined ultrasonographically. Only 60% of horses with histopathologically confirmed GCTs demonstrated elevations in serum inhibin concentrations. Twenty‐five percent of horses had histopathologically confirmed bilateral GCTs. Ninety‐four percent of horses that underwent ovariectomy were discharged from the hospital. Standing laparoscopic ovariectomies had a lower complication rate (34%) than other surgical methods. Long‐term complications were uncommon (11%). In conclusion, histopathologically confirmed GCTs can occur in the presence of normal serum inhibin concentrations. The high prevalence of bilateral GCTs indicates that thorough evaluation of the contralateral ovary is necessary. Surgical ovariectomy for treatment of GCTs has favourable short‐ and long‐term outcomes. Standing laparoscopic ovariectomy may be associated with a lower complication rate than other surgical methods.     

Equine nonneoplastic abnormal ovary in a draft mare with high serum anti-Müllerian hormone: a case study

We performed a standing hand-assisted laparoscopic ovariectomy in a draft mare that presented with high serum anti-Müllerian hormone (AMH) level and had an enlarged single cystic ovary. Histopathological examination revealed no tumor cell proliferation in the ovary, but the presence of a large ovarian cyst was confirmed. In the diagnosis of abnormal ovaries in mares, a comprehensive assessment should be performed, including the monitoring of ovarian morphology and biomarkers over time, to determine the disease prognosis and treatment plan. The case of this mare with a nonneoplastic abnormal ovary and increased serum AMH level was rare. We suggest that standing hand-assisted laparoscopic ovariectomy is useful for the removal of large ovaries in draft mares.     

Low Concentration of Anti-Müllerian Hormone in Mares with Delayed Uterine Clearance

Luteal dysfunction has been observed in mares with defective uterine clearance. Association of low ovarian reserve with luteal dysfunction and abnormal endometrial thickness has been reported in bovine. Anti-Müllerian hormone (AMH) has been indicated as a marker for ovarian reserve in bovine and originates primarily from the ovary in equine. The present study evaluated serum AMH concentration in mares with delayed uterine clearance versus that in mares without delayed uterine clearance. Of 49 mares assigned to the study, 12 and 37 mares were diagnosed with and without delayed uterine clearance, respectively. Delayed uterine clearance was determined based on history and the observation of intrauterine fluid in ultrasonographic examination 24 hours after natural breeding. Serum AMH concentration was measured during estrus. Concentration of AMH was lower in mares with delayed uterine clearance (0.4 ± 0.1 ng/mL) than in those without delayed uterine clearance (1.1 ± 0.1 ng/mL). In conclusion, the present study indicated possible associations between ovary-lined mechanisms and uterine clearance failure in mares.     

Endocrinological characterization of an ovarian sex cord–stromal tumor with a Sertoli cell pattern in a Japanese Black cow

A Japanese Black cow was evaluated for prolonged post‐partum anestrus and enlargement of the right ovary. Transrectal ultrasonography revealed that the right ovary was markedly enlarged and had a solid appearance, while the left ovary was small and inactive. The presumptive diagnosis was directed towards granulosa‐theca cell tumour (GTCT) which was supported by markedly elevated plasma anti‐Müllerian hormone (AMH; 332.0 ng/ml), oestradiol (E₂; 103.3 pg/ml) and immunoreactive inhibin (ir‐INH; 2.1 ng/ml) in comparison with the diagnostic cut‐off points for bovine GTCTs. Since the cow had been infertile and had swelling of the udder, slaughter was chosen. Histopathological examination revealed that the tumour was an ovarian sex cord–stromal tumour (SCST) with a Sertoli cell pattern. These findings suggest that plasma AMH, ir‐INH and E₂ could be possible biomarkers for bovine ovarian SCST with a Sertoli cell pattern, whereas this case could not be distinguished from GTCTs based on endocrinological profile.     

Evaluation of a Motorized Morcellator for Laparoscopic Removal of Granulosa‐Theca Cell Tumors in Standing Mares

Objective: To describe a motorized morcellator technique for laparoscopic removal of granulosa‐theca cell tumors (GCT) in standing mares and to evaluate long‐term outcome. Study Design: Case series. Animals: Mares (n=7) aged 4–15 years, with unilateral GCT. Methods: Tumor size was determined by transrectal palpation and ultrasonography. Standing sedated mares had 3 laparoscopic portal sites in the paralumbar fossa. After laparoscopic observation of the GCT, the mesovarium was desensitized, the ovarian pedicle transected with a LigaSure^® device, and the ovary grasped with forceps and cut in cylindrical tissue blocks using a motorized morcellator. Tissue blocks were removed through the laparoscopic sleeve. Outcome was determined by telephone interview of owners 6–40 months after surgery. Results: Estimated ultrasonographic median GCT diameter was 17 cm (range, 10–22 cm). Surgical time was 2–4.5 hours. There were no surgical complications. Two mares had mild subcutaneous emphysema at the portals after surgery. Convalescence was short, owners were satisfied with cosmetic outcome, and clinical signs associated with GCT did not recur. Conclusion: The motorized morcellator allows piecemeal removal of large GCT through a relatively small laparoscopic portal. Surgical complications were rare and the cosmetic outcome is favorable. Clinical Relevance: A motorized morcellator is a safe and minimally invasive technique for laparoscopic removal of GCT in mares.     

The effects of dexamethasone and prednisolone on pituitary and ovarian function in the mare

Reasons for performing study: Persistent mating induced endometritis is among the most common causes of infertility in the mare. Recently, improved pregnancy rates have been reported when corticosteroids were administered to ‘problem mares’ specifically, to modulate the post mating inflammatory response; however, the effect of treatment on pituitary and ovarian function requires further study. Objectives: To evaluate the effects of prolonged treatment with glucocorticoids on pituitary and ovarian function. Methods: Eighteen cycling Quarter Horse mares in early oestrus were assigned randomly to one of 3 treatment groups: dexamethasone 0.05 mg/kg bwt i.v. twice a day, prednisolone 0.5 mg/kg per os twice a day, or placebo for 5 days. Mares were examined by ultrasound daily to evaluate reproductive function. Blood samples were collected daily to measure luteinising hormone (LH), progesterone and cortisol levels. Results: Dexamethasone treatment caused greater (P<0.05) suppression of endogenous cortisol concentration (9.4 ± 1.1 ng/ml) compared to prednisolone‐ (41.9 ± 4.0 ng/ml) or placebo‐treated mares (32.4 ± 3.8 ng/ml). After 24 h, mares treated with dexamethasone exhibited lower uterine oedema scores than prednisolone‐ or placebo‐treated mares. An ovulation rate of 40% was observed in dexamethasone‐treated mares (2/5) compared to 83% for prednisolone (5/6) and 100% for placebo‐treated (6/6) mares. An absence of a LH surge was noted in 3 of 5 dexamethasone‐treated mares and one of 6 prednisolone‐treated mares. Conclusions: Repeated administration of dexamethasone to mares in oestrus is associated with decreased uterine oedema, suppression of LH and a high rate of ovulation failure. It is recommended that dexamethasone treatment is limited to only 1 or 2 days and that a lower dose is considered in the management of persistent mating induced endometritis to avoid potential adverse affects on reproductive function.     

Double ovarian tumour in the mare: Case report

Ovarian tumours in mares represent 2.5% to 6% of the most frequent neoplasms found in the equine species, with a higher chance of benignity. This study aims to describe a case of two different tumours found in the same ovary of a mare that presented clinical signs of suppressed oestrous cycle during 5 years. After unilateral ovariectomy, the ovary was sent to the histopathology examination which determined a mixed tumour of granulosa cell and leiomyosarcoma. After treatment, the mare returned to oestrus and got pregnant in the next season.     

Sustained-release deslorelin acetate implants disrupt oestrous cyclicity in the mare

There is a need for a safe, effective and practical method of oestrus suppression in the mare. The aim of this study was to monitor ovarian activity in mares exposed to either 9.4 or 28.2 mg deslorelin acetate, a GnRH agonist, in the form of a sustained-release implant. Following oestrus synchronisation, mares were randomly assigned to one of three groups (n = 4 per group) and administered either one (Des1 group; 9.4 mg) or three (Des3 group; 28.2 mg) implants of deslorelin acetate (Suprelorin-12, Virbac Australia) or one blank implant (Control group; Virbac Australia). Mares underwent weekly blood sampling for 12 weeks following implant placement (Day 0–Day 84), with transrectal palpation and ultrasonography of the reproductive tract at all sampling timepoints except Days 56, 70 and 77. All mares showed baseline serum progesterone concentrations (SPC; ≤1.3 nmol/L or 0.4 ng/ml) on Day 0. Cycling Control mares showed typical oestrous cyclicity characterised by peaks and troughs in SPC over time. Four of eight treated mares demonstrated a sustained elevation in SPC after the initial ovulation after implant placement; SPC declined to baseline levels (Des1 group; 2 mares) or remained elevated (Des3 group; 2 mares) at the final sampling timepoint on Day 84. Oestrous cyclicity was erratic in three of the remaining four treated mares. In total, 87.5% (7 of 8) of treated mares showed atypical oestrous cyclicity after implant placement. These results suggest that deslorelin acetate disrupts oestrous cyclicity in the mare, which warrants further research.     

Prolactin,Androstenedione and IGF1 Serum Concentrations During Induced Follicular Growth by eCG Administration in the Bitch

The oestrus cycle in the domestic bitch, a monoestrous species, differs considerably from that of other veterinary domestic animals species. In the bitch the combined use of eCG and hCG is effective to induce oestrus predictably and safely (Stornelli et al., Theriogenology, 78, 2012 and 1056). Although several studies were done to describe the hormonal changes during the canine oestrus cycle, to our knowledge none was done to describe the hormonal changes during induced follicular growth after the administration of eCG. The aim of this work was to study prolactin (PRL), insulin‐like growth factor (IGF1) and androstenedione (ANDR) serum concentrations during follicular growth induced by a single dose of eCG administered to late anoestrous bitches. PRL and ANDR concentrations were lower before than after eCG TRT (before eCG vs pro‐oestrus, oestrus and dioestrus; 4.3 ± 1.8 ng/ml vs 6.5 ± 1.6 ng/ml, p < 0.05; 0.08 ± 0.2 ng/ml vs 0.42 ± 0.16 ng/ml, p < 0.05). Conversely, IGF1 concentrations were similar before and after eCG TRT (286.0 ng/ml ±32.2, p > 0.53). Additionally, PRL concentrations were similar before oestrus compared to during oestrus and dioestrus (6.9 ± 1.7 ng/ml, p > 0.19). Furthermore, IGF1 concentrations were higher before and during oestrus compared to first day of dioestrus (286.1 ± 29.8vs 200.4 ± 29.2 ng/ml, p < 0.01). On the contrary, ANDR concentrations were lower before and during oestrus compared to first day of diestrum (0.35 ± 0.17 ng/ml and 0.38 ± 0.15 vs 0.68 ± 0.17 ng/ml, p < 0.05). These results show that treatment with a single injection of 50 IU/kg of eCG in late anoestrous bitches successfully induced changes in follicular growth which were paralleled with changes in PRL, IGF1 and ANDR serum concentration similar to those occurring during a normally occurring oestrous cycle. In addition, our results suggest that IGF1 in the bitch could play an important role in ovarian folliculogenesis.     

Treatment of haemoperitoneum secondary to ruptured granulosa cell tumours in two mares

This report describes two cases of successful surgical management of granulosa cell tumours (GCT) in mares presenting with haemoperitoneum (HP). Controlled abdominal drainage was initially attempted in Case 1 but was not successful. A ventral midline exploratory laparotomy allowed removal of a haemorrhaging 13 kg GCT. The mare made a full recovery and returned to normal work as a driving pony 11 months post-operatively. In Case 2 controlled abdominal drainage was followed by standing left flank laparoscopic visualisation of the bleeding ovary and transection of the ovarian pedicle by electrocautery. The GCT was then removed via a ventral midline incision due to its large size. Haemoperitoneum can be associated with GCTs and in some cases is severe enough to prompt emergency treatment. Stabilisation of the patient and removal of the haemorrhaging GCT can lead to a successful outcome.     

Attempted conversion of twin to singleton pregnancy in two mares with associated changes in plasma oestrone sulphate concentrations

The removal of one of twin embryos was attempted by infusion of 24% (w/v) saline into the gestation sac in 2 mares by laparotomy. The treatment was successful in one mare (Case 1) and the untreated embryo remained viable. However, neither foetus survived in the second mare (Case 2). Plasma oestrone sulphate (E1S) concentrations fell immediately after treatment in both mares but recovered to approximately 50% of pretreatment levels in Case 1. In Case 2 plasma E1S concentrations declined steadily and were less than 1 ng/ml within 6 days of treatment. These preliminary results suggest that the method may be useful for selective removal of one of twin embryos in mares. Furthermore, plasma E1S concentrations may be a useful indicator of embryonic viability.     

The effect of an extended artificial photoperiod and gonadotrophin-releasing hormone infusions in inducing fertile oestrus in anoestrous mares

The occurrence of fertile oestrus early in the breeding season is of paramount importance to the Thoroughbred industry to facilitate early conception. This paper compares 2 techniques for inducing fertile oestrus in anoestrous mares using either an extended photoperiod alone or together with gonadotrophin-releasing hormone (GnRH) infusions. Eleven mares were placed under conditions of 16 h light and 8 h darkness and 5 of these were implanted with osmotic minipumps delivering approximately 100 ng GnRH/kg/h for 28 days (treated mares). The treated mares ovulated 27.7 days earlier than and conceived 32 days earlier than the 6 mares not given GnRH. GnRH-induced ovulations were followed by a competent luteal phase. The combination of GnRH pumps implanted 2 weeks before commencement of service together with extended photoperiod from July 1 has promise in assisting the stud breeder to improve reproductive efficiency on commercial stud farms.     

Onset of luteal activity in non-foaling mares during the early breeding season in Finland.

The luteal activity in mares was studied in the Equine Research Station (ERS) and in trotting stables (TS) in South-Finland. The mares were Standardbreeds in the TS and mainly Finnhorses in the ERS. Between January and June blood was collected once a week for serum progesterone determinations. The mares in the ERS were distributed in 1 of 3 groups: three-years old not yet in training (N = 38), brood mares (N = 21) and mares in training (N = 47). A 4th group was the mares in training in the trotting stables (N = 73). Every 5th mare in the ERS and every 4th mare in the trotting stables were cycling already at the beginning of the year. Onset of luteal activity in anoestrous mares was most common in the middle of May. Over 95% of the mares were cycling at the beginning of June. In the ERS 40% of the Finnhorse mares in training were cycling through the winter. The three-years old and the brood mares were all anoestrous during winter. They started to cycle on average before the middle of May. Anoestrous training mares started before the middle of April. Anoestrous Finnhorse mares began to cycle later than warm blooded mares in all of the groups studied. Mares which had foaled the previous year were more often anoestrous during the winter than dry mares. The time of year when cycling began in a particular mare tended to be the same from year to year (p less than 0.01).     

The electrosurgical treatment of endometrial cysts in the mare

A minimally invasive technique for the removal of endometrial cysts is described. Intraluminal cysts hinder the migration of the embryo through the uterus in early pregnancy and in a later stage hinder placenta development and hence diminish the chance of successful pregnancy. Cysts can also give rise to false-positive results in early pregnancy tests. Endometrial cysts located in the lumen can be removed surgically from the standing mare. After placement of the endoscope, a wire is placed, via the biopsy channel, around the base of cyst, which is then cut through by cauterization. The cyst is removed with the help of forceps. Five barren mares with multiple cysts were treated in this way during the anoestrous period for October to February. Three of these mares became pregnant during the next oestrus period.     

Effects of oral supplementation with β-carotene on concentrations of β-carotene, vitamin A and α-tocopherol in plasma, colostrum and milk of mares and plasma of their foals and on fertility in mares

In this study, effects of oral β-carotene supplementation to mares (β-carotene group: 1000 mg/day, n = 15; control group: n = 15) from 2 weeks before foaling until 6 weeks thereafter on concentrations of β-carotene, vitamin A and α-tocopherol in plasma, colostrum and milk and plasma of their foals were determined. In addition, effects on fertility were studied. Beta-carotene concentrations increased in plasma and colostrum of β-carotene-supplemented mares compared to control mares (p < 0.05). In mares of both groups, β-carotene concentrations were higher in colostrum than in milk (p < 0.05). In foals, β-carotene concentrations increased with colostrum uptake and were higher in foals born to supplemented mares (p < 0.05; control group: 0.0003 ± 0.0002 μg/ml on day 0, 0.008 ± 0.0023 μg/ml on day 1; β-carotene group: 0.0005 ± 0.0003 μg/ml on day 0, 0.048 ± 0.018 μg/ml on day 1). Concentrations of vitamin A and α-tocopherol were higher in colostrum than in milk (p < 0.05) but did not differ between groups. Concentration of α-tocopherol in plasma of mares decreased over time and in foals, increased markedly within 4 days after birth. All but one mare (control group) showed oestrus within 2 weeks post-partum. Occurrence of oestrus did not differ between groups. More mares of the control group (7/7 vs. 5/12 in the β-carotene group) became pregnant after being bred in first post-partum oestrus (p < 0.05). In conclusion, β-carotene supplementation to mares increased β-carotene concentrations in plasma, colostrum and milk of mares and plasma of their foals but had no positive effects on fertility.