Decidualization of the canine uterus: From early until late gestational in vivo morphological observations, and functional characterization of immortalized canine uterine stromal cell lines.

The apparent lack of classical mechanisms for maternal recognition of pregnancy is one of the most intriguing features of canine reproduction. Consequently, similar levels of circulating luteal steroids are observed in pregnant and non-pregnant dogs. However, the early pre-implantation canine embryo locally modulates uterine responses to its presence, facilitating the successful onset of pregnancy. As a part of this interaction, the canine uterus undergoes a species-specific decidualization. Maternal stroma-derived decidual cells develop, the only cells of the canine placenta expressing progesterone receptor (PGR). There exists an acute need for an in vitro stable cell line model for canine decidualization. Therefore, herein our goal was to establish, immortalize and characterize such a cell line. We immortalized three monolayer dog uterine stromal (DUS) cell lines by stably transfecting them with SV40Tag oncogene. Cells retained their mesenchymal character for over 30 passages, as evidenced by VIMENTIN staining. Genomic incorporation of the SV40Tag protein was confirmed by immunofluorescence and Western blot analyses. Cells submitted to a classical in vitro decidualization protocol (N6,2'-O-dibutyryladenosine-3',5'-cyclic monophosphate) revealed upregulated gene levels of selected major decidualization markers (e.g. PRLR, PGR, IGF1, PTGES). Additionally, the basic decidualization capability of PGE2 was demonstrated, revealing increased levels of, for example, PGR and PRLR gene expression, thereby implying its involvement in the progesterone-dependent decidualization in the canine uterus. In summary, our in vitro model with immortalized DUS cell line could serve as an ideal and unique model to study the underlying molecular and endocrine mechanisms of canine decidualization.

Changes in c-erbB-2 Immunoexpression in Feline Endometrial Adenocarcinomas.

Human epidermal growth factor receptor type 2 (c-erbB-2), an oncoprotein with potential prognostic marker and therapeutic use, is overexpressed in several human and animal tumours. But information regarding this molecule in feline tumours is scarce. This study aimed to assess the changes in the immunohistochemical expression of c-erbB-2 in feline endometrial adenocarcinomas (FEA) compared to normal endometrium. An immunohistochemistry assay using a specific antibody against c-erbB-2 was performed in FEA samples (n = 34) and in normal endometrium in the follicular (FS; n = 12) and luteal (LS; n = 11) stages. In FEA, the c-erbB-2 immunoexpression was assessed in neoplastic epithelial cells whilst in normal endometria it was individually evaluated in the surface and the superficial and deep glandular epithelia (SE, SGE and DGE, respectively). In FS and in LS, all the epithelia were positive for c-erbB-2; positivity was higher in the SE and the SGE than in DGE. Twenty of the 34 FEA samples (58.8%) were positive for c-erbB-2 immunolabelling. Nevertheless, its expression was higher in all the epithelia in the FS compared to FEA (p ≤ 0.0001) or the LS (p = 0.016). The results presented herein suggest that c-erbB-2 molecule is differently expressed in the feline endometrium through the oestrous cycle and though it may also be involved in feline endometrial carcinogenesis, a question remains unanswered on the importance of additional pathways of epithelial proliferation in the neoplastic changes in feline endometrium.

Resident Macrophages and Lymphocytes in the Canine Endometrium.

Resident immune cells play a major role in endometrial immunity and in tissue homoeostasis. This study aimed to analyse the distribution of macrophages, B and T lymphocytes (respectively, Mø, B-Lym and T-Lym) in the canine endometrium throughout the oestrous cycle and in late involution (at the proestrus stage post-parturition). An immunohistochemistry technique was used on samples from 50 post-pubertal healthy female dogs, of which five in late post-partum. The distribution of resident immune cells was analysed in three endometrial layers (superficial, intermediate and basal areas). Mø, B-Lym and T-Lym were demonstrated to reside in the endometrium in all the stages of the canine cycle; their numbers being considerably higher during late involution. T-Lym were scattered in the stroma or amidst the glandular epithelium, constituting the predominant immune cell population in anestrus and proestrus, but decreased in number at all other stages. Endometrial B-Lym remained fairly constant during the canine cycle, although its numbers were higher in late involution. Mø counts were higher during anestrus compared to the other stages, the cells being displaced into the superficial endometrial layer. Mø demonstrated the highest level in late involution samples, forming small aggregates below the surface epithelium. The number of immune cells was not normally distributed, suggesting the influence of individual factors, such as age or parity, not explored herein due to limited sample availability. Still, this study provides important information for the interpretation of endometrial biopsies in dogs and for the understanding of the increased susceptibility to uterine infection during dioestrus found in the bitch.

Reproductive Patterns in the Non-Breeding Season in Asinina de Miranda Jennies.

This study aims to characterize the reproductive patterns in Asinina de Miranda jennies during the non-breeding season. Reproductive activity was surveyed in 12 females, aged between 3 and 18 years old, using ultrasound and teasing with a jack. The animals were monitored from September to April, six in each consecutive year. Of these 12 females, nine showed disruption to the normal pattern of ovarian activity during the non-breeding season. Loss of normal cyclicity included anoestrus (41.7%), silent ovulatory oestrus (25%), and persistence of corpus luteum (8.3%). Only three females maintained a regular cyclic pattern with oestrous behaviour during the non-breeding season. Anoestrus began in early November and lasted for an average of 147 ± 28 days (113-191 days), ending near to the spring equinox. Onset of silent oestrous cycles began more erratically, between October and February. In both groups the first behavioural ovulation of the year occurred around the time of the spring equinox. Disrupted reproductive activity was preceded by a shorter oestrous cycle only in females entering anoestrus. The mean follicle size in the first ovulation of the year was larger than in the reproductive season (44.7 ± 2.45 mm vs 39.2 ± 3.60 mm) in anoestrous jennies with protracted oestrus. Though age and body condition score (BCS) were associated, changes in BCS below a threshold of four points (for anoestrus) and five points (for silent oestrus) contributed greatly to disruption of reproductive cycles. BCS in females with regular oestrous cycles during the winter season remained unchanged or exceeded five points prior to the winter solstice.

Immunohistochemical expression of cyclooxygenase-2 (COX-2) in feline endometrial adenocarcinoma and in normal and hyperplastic endometria.

Cyclooxygenase-2 (COX-2) is overexpressed in several human and animal neoplasms, including the human endometrial carcinoma. It has been suggested as a prognostic marker and a potential therapeutic target. This study aimed to (i) clarify histological aspects of feline endometrial adenocarcinomas (FEA) of the papillary serous type and (ii) characterize COX-2 immunohistochemical expression in normal, hyperplastic and neoplastic endometrium in this species. Archived paraffin-embedded tissue samples of 33 FEA, eight cystic endometrial hyperplasias (CEHs) and 21 samples of normal, healthy endometrium in the follicular (FS; n = 10) and luteal (LS; n = 11) stages were evaluated. Histological evaluation of haematoxylin and eosin-stained sections of the FEA revealed a papillary proliferation of neoplastic cells of serous type, accompanied by clear and multinucleated cells. Other architectural arrangements mainly included solid and tubular growth. Randomly distributed areas of necrosis within the tumours were commonly observed. Invasion of the myometrium, of the serosa and of the vascular and/or lymphatic vessels was not constant features. The mean number of mitoses was higher in FEA compared to non-neoplastic endometrium. COX-2 scores were lower in FEA (p = 0.003) and CEH (p = 0.05) when compared to normal epithelium (NE). The loss of the membrane apical reinforcement in epithelial cells was observed in FEA samples, which was accompanied by the dislocation of COX-2 labelling into the cytoplasm and the perinuclear area; in contrast, in epithelial cells in the healthy and hyperplastic endometria, the immunoreaction showed the characteristic pattern of apical membrane reinforcement, suggestive of the membrane polarization. COX-2 epithelial scores were higher in the FS than in the LS. No differences were found in stromal COX-2 expression between normal, CEH and FEA groups, but it was higher in the LS than in the FS. In summary, loss of COX-2 compartmentalization in neoplastic epithelial cells might be one of the molecular events underlying endometrial carcinogenesis.

Fertility in Adult Bitches Previously Treated with a 4.7 mg Subcutaneous Deslorelin Implant.

The absence of fertility problems in male dogs after a single treatment with deslorelin acetate (Suprelorin(®)) is well acknowledged. However, reports on the application of deslorelin in the bitch and information concerning fertility after implant treatment are still limited. In this retrospective study, data concerning induced and spontaneous oestruses of 39 bitches from 17 breeds, treated with deslorelin acetate implants (4.7 mg Suprelorin(®), Virbac, France), were retrieved to assess post-treatment fertility (ovulation rate, pregnancy rate and litter size). Animals were grouped according to treatment characteristics: group 1 (Gr1) - females submitted to oestrus induction, showing natural oestruses afterwards (n = 19); group 2 (Gr2) - females re-implanted with 4.7 mg deslorelin acetate to re-induce oestrus, showing subsequent spontaneous post-implant oestruses (n = 7); and group 3 (Gr3) - females submitted to a 4.7 mg deslorelin acetate implant for oestrus suppression, evaluated at subsequent spontaneous post-implant oestruses (n = 13). Comparison of fertility traits between induced and post-treatment spontaneous oestruses in Gr1 and Gr2 (short treatments), or between spontaneous oestruses after long-treatment schedules (Gr 3) revealed a slightly better performance in spontaneous cycles compared with induced cycles: ovulation rate post-treatment was 97.1%, 94.1% and 94.4% and the pregnancy rate post-treatment was 91.2%, 88.9% and 84.6% for groups 1, 2 and 3, respectively. Nevertheless, fertility in induced and post-treatment oestruses was considered normal. Moreover, the individual litter size did not differ within groups between induced and spontaneous cycles. From these findings, we concluded that treatment with 4.7 mg deslorelin implants did not compromise the bitches' fertility in subsequent oestruses.

Priapism associated with lumbar stenosis in a dog.

Priapism, a persistent long-lasting involuntary erection of the penis, is uncommon in dogs. In this report, the case of a 13-year-old male Pointer, referred to our services due to persistent exposition of the penis, is described. This condition was consecutive to an intermittent priapism situation lasting for several days, which has been initially attributed to the inflammation and haematoma associated with a perianal bite. The owners became unable to retract the penis into the prepuce. At presentation, the dog was anorectic for 48 h, intolerant to manipulation, and showed poor body condition and unsteady locomotion. During physical evaluation, a marked engorgement of the local vessels in the prepuce and penis was found. An abdominal X-ray was asked under the suspicion of a neurogenic origin for the clinical situation, which showed evidences of spondylosis. After discussion of the clinical condition, the owners asked for euthanasia. The necropsy confirmed the engorgement of the regional vessels deriving from the pudendal arteries and blood accumulation within all the cavernous spaces, accompanied by congestion and thrombosis within the erectile structures of the penis. No significant changes were observed in the pelvic organs that could be at the origin of priapism. The lumbar-sacral spinal regions were carefully inspected and evidenced signs of L7-S1 stenosis due to spondylosis. The case presented herein is a rare situation of priapism of neurogenic origin in a dog. Necropsy findings suggest that it was consecutive to cauda equina compression due to lumbar spinal stenosis.

Feline endometrial adenocarcinoma in females <1 year old: a description of four cases.

Uterine neoplasms of epithelial origin are rare in cats and most often are described in older females. Yet, in less than 2 years, four ovariohysterectomy specimens were submitted from different practices to the Laboratory of Histology and Anatomical Pathology, at UTAD (Vila Real, Portugal), that emitted a diagnosis of feline endometrial adenocarcinoma. Untypically, all the females were aged <1 year old at the surgery. Access to the clinical files was requested to document the clinical features of the four cases, including any complementary data available, to construct the present case reports. The clinical situation developed with discrete signs, but vulvar discharge was present in three cases, ranging from bloody to brownish or colourless, and from purulent to mucous. The females were in dioestrus, although the oestrus remained unperceived in most cases. In this study, the four clinical situations are described and discussed on the basis of available literature, highlighting the aspects that may impair an early diagnosis and that may favour the progression of the disease and also that age should not be an excluding criteria when analysing the differential diagnosis list.

Uterine prolapse with associated rupture in a Podengo bitch.

A case of uterine prolapse coexisting with uterine horn rupture in a 3-year-old Portuguese Podengo bitch, which is an uncommon occurrence, is described. The female was presented with a history of recent parturition, with delivery of four healthy puppies that were normally tended and nursed. The situation developed after an uneventfully pregnancy, and no direct causative factor was identified. The duration of the prolapse was unknown, but considered to be recent because of the swollen reddish appearance of the tubular everted mass. No foetus was found in the uterus or the abdominal cavity. The female was presented in good physical condition, without signs of shock or haemorrhage. During surgical treatment, the uterus was replaced to its normal position followed by ovary-hysterectomy at 12 h from admittance.

Tumour necrosis factor in the canine endometrium: an immunohistochemical study.

Tumour necrosis factor (TNF), a pleiotropic cytokine that regulates cell growth and differentiation as well as the synthesis of other cytokines, has been identified in the uterus of several species describing a cyclic pattern, eventually under ovarian steroid regulation. Information is yet limited on the presence of TNF protein in the canine endometrium during the oestrous cycle and early pregnancy. This study depicts the temporal immunolocalization of TNF in the bitch endometrium along the oestrous cycle and changes associated with the early steps of embryo invasion. TNF immunolabelling was found in both the stromal fibroblasts and epithelial components of the canine endometrium in all stages studied. Stromal immunostaining was more intense than that of the epithelia, in all the stages of the oestrous cycle. In addition, a tendency for a decrease in the surface epithelium intensity score was found in early dioestrus. A positive glandular content was only observed in anoestrus and proestrus stages. In early pregnancy (days 13-16), TNF immunolabelling was detected at the embryo-maternal surface, in the syncytium cords and the trophoblast, as well in the endometrial stroma and the basal endometrial glands, but not in the lacunar epithelium. The overall TNF immunoreactivity was higher in early pregnancy samples in comparison with those of the early dioestrus and dioestrus stages, suggesting it plays a role during implantation.

Cervical hypoplasia in jennets: a description of two cases.

The authors describe two cases of cervical hypoplasia in jennets, detected during the gynaecological evaluation of two purebred females of the Portuguese donkey breed Asinina de Miranda. These jennets aged 10 and 15 years were referred for consultation by the local breeders association because of infertility detected during an artificial breeding programme. In both jennets the cervix was small, shorter than the usual and closed. In one of the females, outflow of semen during artificial insemination was reported. Clinical and ultrasonographic evaluation of the genital tract demonstrated that both ovaries were normally sized in both animals and presenting structures suggestive of regular ovarian activity. In one of the jennets, a small amount of air in the uterus was evidenced. In the other, signs indicative of atresia of the cervical canal were found. Vaginal examination showed that the cervix was reduced in size and shallowed in both animals. A diagnosis of congenital cervical hypoplasia was made in both cases and the animals were excluded from reproduction.

Histopathologic and immunohistochemical exam in one case of canine endometrial adenocarcinoma.

Canine endometrial carcinomas are rare, and mostly occur in geriatric bitches. In this work, the uterus of a 10-year-old female Boxer evidencing an endometrial carcinoma on the body of the uterus was used to describe the histopathological features of the tumour and to study its immunophenotype. In this work, a panel of immunomarkers (cytokeratins AE1/AE3 and 14, vimentin, CD10 and Ki-67) was applied to the endometrial carcinoma to establish the staining patterns indicative of the tumour agressiveness and cellular differentiation. Additionally DNA ploidy was also performed. In this case, the tumour showed papillar pattern, with large pleomorphic, anaplastic cells and also some aberrant multinucleated and giant cells. In some areas of the tumour, it was also observed cytotrophoblastic-like cells outlining the papillae. Cytokeratin AE1/AE3 expression was detected in the luminal neoplasic cells. Cytokeratin 14 positivity was sporadic and irregular, and was observed mainly in the luminal epithelium. Only stromal and aberrant cells showed a positive staining to vimentin. Positive membranous staining to CD10 was evidenced by clear epithelial, cytotrophoblastic-like cells at the tumour surface but not by the stromal cells. The mitotic and Ki-67 indices were low, suggestive of a weak aggressiveness of the tumour. The multinucleated and giant cells evidenced a positive immunostaining to CK AE1/AE3, and CD 10; its positivity to vimentin was sporadic. This study aims to contribute to the advancement of the knowledge in canine endometrial carcinoma immunophenotype.

Priapism secondary to perineal abscess in a dog - a case report.

A 7-year-old intact male Boxer was referred to our services at the Veterinary Teaching Hospital of the University of Trás-os-Montes and Alto Douro, suffering from a persistently erect penis (including the bulbus glandis) that had been exposed for several days. Radiographic and ultrasonographic examinations detected a 5.0 x 3.5 cm mass located dorso-laterally to the urinary bladder. The microbial culture of the mass revealed Staphylococcus spp. At that time, we suspected the involvement of an abscess in the origin of the priapism. Medical and surgical treatments were promptly instituted, which allowed for penile withdrawal into the prepuce; however, the resolution of the penile erection was not accomplished in the following days and penile amputation was required. Histological evaluation of the excised penis revealed extensive infarction of the erectile tissue of the pars longa and bulbus glandis, and also of the blood vessels of the penis. Following penile amputation and antimicrobial therapy, the animal fully recovered. Ultimately, the animal died as a consequence of gastric torsion. At necropsy, some lesions compatible with a previous perforation of the intestinal wall were recorded. The data gathered from the anamnesis, the physical and imaging examinations, along with the post-mortem findings, allowed us to conclude that in this clinical case the primary cause of priapism was a perineal abscess due to bowel perforation.

A complex intersex condition in a Holstein calf.

A case of disrupted embryonic development of the genital tract in a newborn Holstein calf is described. The physical examination of the calf evidenced several abnormalities, like atresia ani, rudimentary external genitalia and caudal vertebral agenesis. On necropsy, the excised genitalia consisted of bilateral streak gonads, apparently normal uterine tubes, a fluid-filled uterus, a long vagina and a very narrow clitoris-like structure covered with a discrete skin-fold. The urinary tract seemed normal and the urethra's opening was at the vestibule-vaginal junction. A cytogenetic analysis was requested. Karyotype revealed the existence of Y chromosome material in the two X chromosomes. However, the search for the sex-determining region Y (SRY) showed that this was an apparently absent gene. The histological examination of the gonads revealed the existence of ovarian dysplasia. Uterine sections evidenced the absence of the uterine epithelium, with only sporadic caruncles. Under microscopic examination, the uterine tubes and vagina structure was normal. The external genitalia sections revealed the existence of a skin-fold covering an erectile structure surrounding the urethra, a structure more similar to a penis than to a clitoris. This is an unusual situation of gonadal dysplasia combined with genital tract anomalies in cattle, probably associated to a genetic defect.

Multioocyte follicles in domestic dogs: a survey of frequency of occurrence.

Multioocyte follicles (MOFs) or polyovular follicles have been reported infrequent in the ovaries of the bitch, decreasing with the age. In this study, the routine observation of the ovaries allowed to verify that the existence of MOFs was far more frequent than previously reported. Ovaries from 150 genitalia, excised during ovariohysterectomy of bitches of different breeds and ages were used. The mean prevalence of MOFs was 40.7%, and the prevalence was higher in young animals (68.4 and 62.2%, in prepubertal and in bitches under 1 year, respectively). In 7-8 years old bitches, the occurrence decreased to 30.4%, and it decreased again to 14.3% in 10 or more years old bitches. It was also more frequent in mongrels than in pure-breeds (52.3-25.5%, respectively). Most follicles contained 2-3 oocytes, but follicles containing up to 10 oocytes were also observed. When the number of oocytes was higher than 3, oocytes of various morphological appearances could be noticed within the follicle. These observations show that the presence of MOFs can affect the number of oocytes recovered in assisted reproductive protocols and may also influence the ovulation rate and prolificity of these animals.

Cysticercus tenuicollis vesicle in fetal structures: report of a case.

Cysticercus tenuicollis is the larval stage of the canine tapeworm Taenia hydatigena, the presence of which has been reported in wild and domestic ruminants all over the world. It is a common parasite of small ruminants in the north of Portugal. C. tenuicollis is generally seen attached to the omenta, the mesenteries or also found in the liver. In the ewe, tissue lesions have been associated with degenerative cysts or with oncosphere migrations. Unusual locations of the cysticerci of T. hydatigena have been described. The most frequent unusual locations are in the lungs, the kidneys and the brain. Less common locations have been reported to occur in the ovaries, uterine tubes, uterus, cervix and vagina. In the case described here, and for the first time, an aberrant location of a C. tenuicollis vesicle was found inside the chorion-allantoic membrane of a goat's foetus, in a gemelar gestation of approximately 70 days. Finding a C. tenuicollis vesicle inside fetal membranes forewarns of the possibility of larval migrations into the fetal structures during pregnancy, which is particularly concerning in human populations that are infested.

Early embryo development, number, quality, and location and the relationship with plasma progesterone in dogs.

This research assessed the developmental stages and morphological quality of dog embryos collected during different stages of pregnancy as well as the relationship with serum progesterone recorded at insemination and embryo collection. Embryos were collected from 23 young mature bitches, that had been inseminated with fresh semen 3-6 days after the LH surge (day 0). Embryo flushing was performed on pregnancy days 8-11 (Group 1), 12-15 (Group 2), or 16-20 (Group 3). The location, number and morphological characteristics of the embryos were evaluated. A total of 120 embryos and 25 unfertilized oocytes were collected from bitches with a total of 156 corpora lutea (CL). The mean total embryo yield (total of embryos/CL) was 76.7 ± 5.9%, and the mean embryo recovery rate (number of flushed embryos/number of CL) was 70.6 ± 6.6%. The mean ovulation rate was 6.8 ± 0.5 and the mean number of embryos per bitch was 5.2 ± 0.6. Oocyte fertilization occurred following oocyte maturation. Most embryos in Group 1 (70.0 ± 18.6%) were collected at the 2 to 16 cell stage. The morula stage was first observed on day 11. Expanded blastocysts (EBLs) and hatched blastocysts were first flushed from the uterus on days 13 and 14, respectively. The EBL was the most abundant stage in Groups 2 and 3. After day 19, some embryos (n = 8) had already adhered to the endometrium. Although most recovered embryos were classified as very good, a greater number of low quality embryos was collected in the later gestational periods. A significant variation in the embryonic stages and location of embryos in early canine pregnancy was observed, as embryos entered the uterus independently of their developmental stage. Embryo yield and quality were independent of the serum progesterone concentration at insemination and recovery.

A case of blind vagina in a crossbred Lusitano mare.

This report describes a case of blind vagina diagnosed during a pre-breeding evaluation of a 5-year-old crossbred Lusitano mare. This mare was mated twice during the previous breeding season but remained open. Clinical evaluation revealed the existence of follicles in the ovaries and an enlarged uterus. An ultrasound examination showed that a granular free-floating fluid distended the uterus. No connection was detected between the uterus and the vagina and a presumptive diagnosis of congenital vaginal obstruction was raised. On vaginal examination it was noticed that the vagina was short in depth and ended as a blind pouch. The typical cervical morphology was not observed. Careful evaluation of the clinical situation, both during the follicular and the luteal phases of the mare's oestrous cycle, lead to a diagnosis of segmental aplasia of the cranial vagina. The mare had a normal karyotype on cytogenetic examination [64XX].

Immunolocalization of E-cadherin and ß-catenin in the cyclic and early pregnant canine endometrium.

Putative changes in E-cadherin and ß-catenin during implantation in dogs are of interest to study, as they are relevant proteins for epithelial integrity. E-cadherin and ß-catenin were immunolocalized in the canine endometrium during the estrous cycle and early pregnancy, using monoclonal antibodies. Both proteins were detected in all types of endometrial epithelia (surface epithelium [SE], superficial glandular, and deep glandular epithelia) at all stages of the estrous cycle and in early placental structures. E-cadherin depicted a gradient of intensity apparently being lowest in the SE to progressively increase toward the deepness of the endometrial glands, regardless of the stage of estrous cycle. The overall immunostaining was, however, weaker at diestrus. In pregnant samples, the trophoblast was conspicuously immunolabeled compared with the endometrial surface lining epithelium. In the latter, the cytoplasmic pattern predominated over the membrane-bound, as was also seen in the decidual cells of the placental labyrinth. In the early placenta, only trophoblast cells and lacunae retained membrane signals. ß-Catenin membrane labeling appeared relatively constant throughout the cycle, although a tendency toward a decrease in intensity was detected at the secretory stages. In addition, a dislocation of the immunoreaction from membrane to the cytoplasm was observed in both the SE and the glandular epithelia at particular stages of the cycle. In early pregnancy, a loss of the membranous pattern was observed in the SE and labyrinth, but neither on trophoblast nor in lacunae. The results show the existence of a softening of the adherens junctional complex in progestagen-dominated stages favoring embryo-maternal interactions and endometrial invasion during canine implantation.

Characterization of the estrous cycle of Asinina de Miranda jennies (Equus asinus).

This study aims to characterize the estrous cycle of Asinina de Miranda jennies in the breeding season, on the basis of data collected from serial ultrasonographic examination and serum progesterone determinations in 14 females during a total of 33 cycles. The length of the interovulatory interval was 23.8 ± 0.55 days, the diestrus and estrus lasting 17.9 ± 0.46 days and 6.65 ± 0.30 days, respectively. Age and body condition score (BCS) affected the length of the interovulatory intervals; BCS also influenced the diestrus length and the time in heat after ovulation (P > 0.05). The incidence of single, double, and triple ovulations was 57.58%, 36.36%, and 6.06%, respectively. Multiple ovulations affected neither the length of the interovulatory interval nor the individual cycle stages (P > 0.05) but lengthened the interval from the beginning of estrus to the last ovulation (P = 0.01). When combined with age, higher BCS affected the ovulation rate (P = 0.001). Deviation of the dominant follicle occurred around Day 8.7 (Day 0 = ovulation) when both single and multiple ovulations were considered. The dominant follicle was larger at divergence in single ovulators (19.18 ± 0.97 mm) compared with that in multiple ovulators (18.05 ± 1.16 mm). The overall maximum follicular diameter before ovulation was smaller in multiple ovulatory cycles than that in single ovulatory cycles (37.2 ± 0.83 mm vs. 40.2 ± 1.41 mm, respectively; P = 0.03). The daily growth rate of dominant follicles was independent of the ovulation rate (P > 0.05) for the intervals before and after the estrus onset. The dominant follicle size and the follicle growth rates were independent of BCS (P > 0.05). Data collected in this study revealed resemblances between Mirandese and other Iberian and Brazilian breeds with regard to estrous cycle characteristics.