A comparison of the uterine proteome of mares in oestrus and dioestrus

Proteomic analysis of mare uterine flush fluid provides a minimally invasive technique for studying protein changes associated with the oestrous cycle. The aim of this study was to identify differentially abundant proteins in the uterine flush fluid of mares in oestrus and dioestrus. In this study, uterine flush fluid samples were collected from eight reproductively healthy mares in either oestrus (n = 5) or dioestrus (n = 3). Proteomic analysis was performed using liquid chromatography‐tandem mass spectrometry. Of 172 proteins identified, six proteins (immunoglobulin lambda‐like polypeptide 1, haemoglobin subunit alpha, alpha‐1B‐glycoprotein, serotransferrin, apolipoprotein A‐1, and haemoglobin subunit beta) were significantly more abundant in oestrus. These proteins may contribute to the endometrial defence system through roles in inflammation, immunity or antimicrobial activity. In other species, some of these proteins have been described as immunoglobulins, negative acute phase proteins or defence agents against micro‐organisms. During dioestrus, immunoglobulin alpha‐1 chain C region‐related, complement factor I, CD 109 antigen and uterocalin, were significantly more abundant. Research in other species suggests that these four proteins contribute to the immune response through proposed immunoregulatory characteristics, complement system involvement or roles in B cell–T cell interactions. In conclusion, ten differentially abundant proteins were identified in the uterine flush fluid of mares in oestrus and dioestrus. Targeted studies on these proteins could elucidate their role in uterine defence mechanisms during the oestrous cycle in the mare.     

Uterine prolapse in a mare leading to metritis,systemic inflammatory response syndrome,septic shock and death

This Case Report describes severe complications associated with uterine prolapse in a mare. A 6‐year‐old Trakehner mare was examined for depression, moderate pain and vaginal discharge 3 days after correction of a uterine prolapse. The clinical examination and haematology revealed that the mare had an infection with systemic inflammatory response syndrome and shock. Due to the uncontrollable, persistent pain, an exploratory celiotomy was performed which revealed severe metritis. During anaesthesia, the mare developed severe cardiovascular compromise and died in recovery. In previously reported cases of uterine prolapse in the mare, the authors warn of uterine injury, broad ligament haemorrhage, metritis, endotoxaemia and laminitis but often have a successful outcome with conventional therapy. This case describes a mare that developed severe complications and death after uterine prolapse. Mares with uterine prolapse require appropriate treatment and vigilant monitoring post treatment to prevent life threatening complications.     

Removal by laparoscopic partial ovariohysterectomy of a uterine leiomyoma assumed to have caused fetal death in a mare

A 13‐year‐old pluriparous Dutch Warmblood mare presented to Utrecht University's Department of Equine Sciences 4 weeks after suspected abortion at 3.5 months gestation, to investigate the nature of a uterine mass and persistent vulval discharge. Transrectal ultrasonographic examination revealed copious flocculent fluid and fetal remnants within the uterus and a 5–6 cm heterogenous mass in the uterine wall at the tip of the right horn. Expulsion of fetal parts and resolution of the coexisting endometritis were effected by a combination of repeated PGF_2a analogue injections to induce oestrus, application of PGE₂ gel to aid cervical relaxation, and daily uterine lavage and antibiotic instillation. The presence of the mass in the uterine wall was confirmed by hysteroscopy and the suspected tumour subsequently removed by partial laparoscopic ovariohysterectomy under standing sedation and local anaesthesia. The histological appearance of the tumour was consistent with a leiomyoma or moderately malignant leiomyosarcoma. Although a follow‐up examination 6 months post surgery revealed uncomplicated healing of the uterus, the owner decided to retire the mare from breeding. Uterine neoplasia is an extremely unusual cause of fetal death in the mare but, in the present case, laparoscopic partial ovariohysterectomy proved a promising, minimally invasive technique for salvaging sufficient uterus to make subsequent breeding a realistic proposition.     

Intra-abdominal hemorrhage associated with a granulosa-thecal cell neoplasm in a mare

A 9-year-old American Saddlebred mare was referred because of abdominal distention and signs of abdominal pain. Copious peritoneal fluid obtained by abdominocentesis appeared to be frank blood. Rectal and ultrasonographic evaluation of the abdomen revealed a large mass at the distal tip of the right uterine horn. The mare was euthanatized and necropsied and the mass was determined to be a granulosa-thecal cell neoplasm. The most common clinical sign of granulosa-thecal cell neoplasm is infertility or abnormal sexual behavior. Hemoperitoneum is infrequently associated with neoplasms in horses.     

Diagnosis of a uterine leiomyoma using hysteroscopy and a partial ovariohysterectomy in a mare

A 14-year-old quarter horse mare had a 2-year history of infertility and the presence of a mass in the right uterine horn. Prior to surgery, the mass was determined to be invasive by using ultrasonography and hysteroscopy. A partial ovariohysterectomy was performed. The mass was diagnosed histologically as a uterine leiomyoma.     

Multicentric T‐cell lymphoma presenting as inferior palpebral swelling in a Standardbred mare

A 6‐year‐old Standardbred mare that presented with bilateral palpebral swelling and a left corneal ulcer was diagnosed with a multicentric T‐cell lymphoma. Other clinical findings included submandibular lymphadenopathy and oedema, pharyngeal oedema and rhinitis. Prior to euthanasia the horse developed keratomalacia, the onset of which was coincidental with the use of high doses of topical and systemic corticosteroids. Although an unusual presentation, palpebral swelling should be considered as a clinical sign of lymphoma.     

Chronic intermittent colic in a mare attributed to uterine marbles

A 13‐year‐old Quarter Horse mare presented for evaluation of chronic intermittent colic. Following extensive diagnostics, abdominal radiographs revealed two round, radiopaque objects in the caudal abdomen. Palpation per rectum and transrectal ultrasonography of the reproductive tract confirmed that the round objects were uterine marbles. Dinoprost tromethamine (Lutalyse, 5 mg i.m. q. 24 h for 2 days) was administered to bring the mare into oestrus, and both uterine marbles were manually removed from the uterus following digital dilation of the relaxed cervix. Follow‐up with the owner 12 months after discharge revealed that the mare had shown no further signs of abdominal discomfort since having the uterine marbles removed. To the authors' knowledge, this is the first published report of chronic intermittent colic attributed to uterine marbles in a mare.     

Pelvic lymphoma as a cause of urethral compression in a mare

A 6-year-old mare was presented for evaluation of dysuria and pollakiuria. A pelvic mass, which caused obstruction of urinary outflow, was identified. Evidence of renal failure was also present. Due to poor prognosis the mare was euthanized. A postmortem examination revealed a pelvic lymphoma that surrounded and compressed the urethra.     

Extreme lymphocytosis with myelomonocytic morphology in a horse with diffuse large B‐cell lymphoma

An 11‐year‐old, 443‐kg Haflinger mare was presented to the North Carolina State University Veterinary Teaching Hospital with a 2‐week history of lethargy and a 3‐day duration of anorexia, pyrexia, tachycardia, and ventral edema. Severe pitting edema, peripheral lymphadenopathy, and a caudal abdominal mass were noted on physical examination. An extreme leukocytosis (154.3 × 10³/μL) and microscopic hematologic findings suggestive of myelomonocytic leukemia were observed. Serum protein electrophoresis revealed a monoclonal gammopathy and urine protein electrophoresis revealed a monoclonal light chain proteinuria. Necropsy and histopathology confirmed widespread neoplastic infiltration in many organs with a heterogenous population of cells; there was no apparent evidence of bone marrow involvement. Immunohistochemistry confirmed presence of a majority of B cells with a limited antigen expression, admixed with a lower number of T cells. Molecular clonality analysis of IgH2, IgH3, and kappa‐deleting element (KDE, B cell) on whole blood and KDE on infiltrated tissues revealed clonal rearrangements, and the KDE intron clones that amplified in blood and in infiltrated tissue were identical. In contrast, the clonality analysis of T‐cell receptor γ revealed no clonality on blood cells and infiltrated tissues. In conjunction with the histopathologic changes, the lesion was interpreted to be composed of neoplastic B cells with a reactive T‐cell population. Polymerase chain reaction testing for equine herpes virus 5 was negative. The final diagnosis was diffuse large B‐cell lymphoma with a marked hematogenous component.     

Treatment of a pulmonary granular cell tumour by repeated transendoscopic diode laser photoablation

A 27‐year‐old Shetland mare was examined for persistent coughing, increased respiratory rate and mild exercise intolerance. Over the previous 5 months, the mare had experienced a persistent cough nonresponsive to medical and environmental management. Upon endoscopic examination of the lower airways, a mass was seen within the left mainstem bronchus; biopsy identified a granular cell tumour. The tumour was successfully reduced by repeated transendoscopic diode laser photoablation over a period of 4 months with no evidence of recurrence at 10 months post treatment.     

Use of a hand-assisted laparoscopic technique for removal of a uterine leiomyoma in a standing mare

A 20-year-old Thoroughbred mare was evaluated because of a 2-year history of infertility. The mare had normal estrous cycles and had been bred 7 times by different stallions. Ultrasonographic examination revealed a homogeneous hyperechoic intramural mass in the tip of the right uterine horn; the mass was also detected via hysteroscopy Unilateral ovariectomy and partial hysterectomy were performed by use of a hand-assisted laparoscopic technique. Leiomyoma was diagnosed via histologic examination of the mass. Unilateral ovariectomy and partial hysterectomy are recommended in mares with leiomyoma in a uterine horn, especially if the tumor is associated with infertility. The hand-assisted laparoscopic technique allows direct visualization of abdominal structures and accurate placement of ligatures without applying tension on the broad ligament, and eliminates the risks and costs of general anesthesia.     

A tumor-related lymphoid progenitor population supports hierarchical tumor organization in canine B-cell lymphoma

Background: Tumors have heterogeneous properties, which could be explained by the existence of hierarchically and biologically distinct tumor cells such as tumor‐initiating cells (TICs). This model is clinically important, as TICs are promising targets for cancer therapies. However, TICs in spontaneous B‐cell lymphoma have not been conclusively identified. Hypothesis/Objectives: Tumor cells with a progenitor phenotype exist in B‐cell lymphoma, reflecting a hierarchical organization. Animals: Twenty‐eight client‐owned dogs with previously untreated B‐cell lymphoma and 6 healthy dogs. Methods: This was a prospective study. Flow cytometry was used to identify lymphoid progenitor cells (LPCs) that coexpressed hematopoietic progenitor antigens CD34, CD117, and CD133, with lymphoid differentiation markers CD21 and/or CD22 in B‐cell lymphoma. The polymerase chain reaction for antigen receptor rearrangements was used to analyze clonality and relatedness of tumor populations. A xenograft model with NOD/SCID/IL‐2Rγ^?/? mice was adapted to expand and serially transplant primary canine B‐cell lymphoma. Results: LPCs were expanded in lymph nodes from 28 dogs with B‐cell lymphoma compared with 6 healthy dogs (P= .0022). LPCs contained a clonal antigen receptor gene rearrangement identical to that of the bulk of tumor cells. Canine B‐cell lymphoma xenografts in recipient mice that maintained LPCs in the tumors were recurrently observed. Conclusions and Clinical Importance: These results suggest the presence of a hierarchy of tumor cells in B‐cell lymphoma as has been demonstrated in other cancers. These findings have the potential to impact not only the understanding of lymphoma pathogenesis but also the development of lymphoma therapies by providing novel targets for therapy.     

Ascites in a 13‐year‐old miniature horse mare with an ovarian granulosa theca‐cell tumour: A manifestation of Meigs' syndrome?

A 13‐year‐old miniature horse mare presented for evaluation of ascites. Abdominal ultrasound showed severe free peritoneal fluid accumulation as well as a caudal abdominal mass. The peritoneal fluid was characterised as a transudate with no evidence of sepsis. Surgical exploration of the abdomen was performed; copious free peritoneal fluid and a large right ovarian mass were discovered. The abnormal ovary was removed, and the mass was determined histologically to be a benign granulosa theca‐cell tumour. Although there was mild peritoneal fluid accumulation in the immediate post operative period, the ascites effectively resolved with removal of the tumour, and the mare returned to a normal lifestyle. This is similar to reports of Meigs' syndrome in human women, in which a benign ovarian mass of significant size causes ascites that resolves once the mass is removed.     

Standing laparoscopic-assisted vaginal ovariohysterectomy in a mare

A 20-year-old Arabian mare presented to Washington State University Veterinary Teaching Hospital for evaluation of haemorrhagic vaginal discharge of 3 months' duration. The referring veterinarian had identified a mass within the uterine wall via transrectal ultrasonographic examination. On presentation, the mare had an unremarkable physical examination with the exception of a mild haemorrhagic vaginal discharge. Rectal palpation was performed and an approximately 9 cm diameter mass with a granular texture was identified associated within the uterine body and left uterine horn. Endoscopic examination of the reproductive tract revealed a linear defect in the ventral uterine wall near the cervix with direct communication into the abdomen. Standing laparoscopic-assisted vaginal ovariohysterectomy (OHE) was performed, which involved laparoscopic facilitated dissection and haemostasis of uterine and ovarian structures, and inversion of the uterus through the cervix. Removal of the uterus was performed vaginally. No post operative complications were noted. Standing laparoscopic-assisted vaginal OHE is an alternative to traditional OHE techniques. This technique allowed for excellent direct visual observation during dissection and ligation and did not require general anaesthesia.     

Disseminated lymphoma with large granular lymphocyte morphology diagnosed in a horse via abdominal fluid and transtracheal wash cytology

A 22‐year‐old Tennessee Walking Horse mare was presented to the Auburn University Large Animal Teaching Hospital with a 3‐day history of lethargy, anorexia, and mild signs of colic. The mare had a several‐month history of weight loss and refractory cough. Physical examination revealed an increased respiratory rate, and crackles and wheezes were heard on thoracic auscultation. Thoracic ultrasonographic examination showed disseminated, minor, bilateral comet tail‐like lesions on the parietal pleural surfaces. Abdominal ultrasonographic examination was unremarkable. Trans‐rectal palpation revealed a firm small colon impaction with concomitant diarrhea. Laboratory data were characterized by a very pronounced acute inflammatory leukogram with severe neutropenia and significant left shift, evidence of hepatocellular damage/necrosis, cholestasis, and possibly mixed metabolic alkalosis and acidosis. On cytologic evaluation of a peritoneal fluid sample, there were many large granular lymphocytes (LGL). Large numbers of LGL were also observed on cytologic examination of a subsequent transtracheal wash. The final cytologic interpretation was disseminated lymphoma with LGL morphology. Due to worsening of the clinical signs and poor prognosis, the mare was euthanized. On necropsy and in histopathologic examination, disseminated lymphoma with LGL morphology was noted in a mesenteric lymph node, lungs, liver, spleen, kidneys, and right dorsal colon. Lymphoma with LGL morphology is rarely diagnosed in the horse. This report provides unique cytologic findings of a case of disseminated lymphoma with LGL morphology in a horse, confirmed with histopathologic evaluation.     

24-Hour Secretion Patterns of Plasma Oestradiol 17β in Pony Mares in Late Gestation

The mare exhibits nocturnal uterine contractions in the last 6 days of gestation. It is hypothesized that estradiol 17β (O17β) may be associated with the nightly increase in uterine contractions. The 24‐h secretion pattern of plasma O17β was measured in 3 pony mares in late gestation to identify changes in release as the mare neared parturition. Blood was collected weekly at 08:00 hours beginning on day 240 and every third day from day 330 until delivery. Serial blood samples were collected from each mare every 30‐min for 24‐h beginning on gestation day 310 and every sixth day thereafter until parturition. Concentrations of O17β were elevated at night with lowest concentrations occurring directly before sunset (p < 0.01). The natural log of the variance was increased at sunset (p < 0.01) and was decreased during the 6‐h period immediately after sunrise. This pattern was especially evident in the 6 days that preceded parturition. The contrast between nocturnal and daytime concentrations of O17β in the last 6 days of gestation may contribute to night‐time delivery in the mare.     

Canine indolent and aggressive lymphoma: clinical spectrum with histologic correlation

Sixty‐three dogs with newly diagnosed lymphoma underwent complete staging and received the same chemotherapy. Diffuse large B‐cell lymphoma was the leading histotype (44.4%), followed by peripheral T‐cell lymphoma (20.6%). Indolent lymphomas accounted for 30.2% of cases. Most dogs with aggressive B‐cell lymphoma had stage IV disease. Dogs with indolent and aggressive T‐cell lymphoma had more often stage V disease and were symptomatic. Liver and bone marrow were predominantly involved in B‐cell and T‐cell lymphoma, respectively. The clinical stage was significantly related to substage, sex and total lactic dehydrogenase (LDH) levels. Aggressive B‐cell lymphomas were more likely to achieve remission. Median survival was 55 days for aggressive and indolent T‐cell lymphoma, 200 and 256 days for indolent and aggressive B‐cell lymphoma, respectively. The prognosis of advanced indolent lymphoma does not appear to be appreciably different from that of aggressive disease. Familiarity with the various histotypes is critical to make the correct diagnosis and drive therapy.     

Prospective evaluation of flow cytometric characteristics,histopathologic diagnosis and clinical outcome in dogs with naïve B‐cell lymphoma treated with a 19‐week CHOP protocol

Canine B‐cell lymphoma is a clinically heterogenous disease; however, it is generally treated as a single disease entity. The purpose of this clinical trial was to prospectively evaluate naïve canine B‐cell lymphoma patients using histopathology, flow cytometry (FC) and a standardized chemotherapy protocol to better define subsets of this disease that may respond differently to treatment. Sixty‐four dogs with naïve multicentric B‐cell lymphoma were treated with a standardized 19‐week CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) chemotherapy protocol. Most of the dogs (84.3%) were diagnosed with diffuse large B‐cell lymphoma (DLBCL), followed by nodal marginal zone (7.8%), small B‐cell (4.7%), Burkitt‐like (1.6%) and follicular lymphoma (1.6%). FC confirmed the diagnosis of B‐cell lymphoma in all cases. There were no clear phenotyping differences between the subtypes of B‐cell lymphoma detectable by our FC panel. The histologic subtypes in this study exhibited a range of forward scatter values on flow cytometry, but all of the DLBCL cases were higher than a value of 469, while the only cases with a lower forward scatter value were follicular lymphoma and diffuse small B‐cell lymphoma. Dogs with DLBCL had a significantly better objective response rate to the CHOP protocol (96.3%) than the non‐DLBCL subtypes (70%, P = .024). The median progression‐free survival time for patients with DLBCL (233 days) was significantly longer than that of all other histopathologic subgroups combined (163 days, P = .0005).