Periparturient Arterial Rupture in Mares: A Postmortem Study

There are limited reports characterizing the occurrence of rupture of uterine and other arteries associated with pregnancy in mares. Cases were retrieved from the records of horses submitted for necropsy to the University of Kentucky Veterinary Diagnostic Laboratory over a 15-year period (1993-2007). Five hundred thirteen horses were diagnosed with periparturient arterial rupture, representing 1.5% of the total equine necropsy cases. Arterial rupture was diagnosed predominately in older mares, with 78% of the horses being ≥15 years of age. Ruptures occurred most commonly during parturition but also were diagnosed before and after parturition. Additionally, during a consecutive 3-year period within this time interval, an extensive workup of cases was performed. In these 71 cases, the area of the arterial rupture was carefully dissected, and the location, size, and appearance of the arterial tear were determined. The uterine artery was the most commonly affected vessel. Although there was overlap, mares with uterine artery rupture tended to be older than those with rupture of other arteries. There was no predilection for the left or right sides, and the uterine artery rupture occurred most often in the first 20 cm from the origin.     

Ventrocaudal cystorrhaphy without urethral sphincterotomy in two broodmares

Two broodmares were diagnosed with rupture of the urinary bladder. One mare revealed abnormalities post partum and the other associated with ante partum uterine torsion. The clinical symptoms included mild abdominal pain, anorexia, decreased urinary volume and increased peritoneal fluid. In one mare, based on the creatinine level of the peritoneal fluid and serum biochemical abnormalities, uroperitoneum was diagnosed. In the other mare, the bladder rupture was found during the celiotomy for surgical repair of uterine torsion that was diagnosed upon rectal examination. Surgery was performed without a urethral sphincterotomy. The vaginal floor was incised in a standing position and the bladder was diverted into the vagina in order to suture the tears located in the ventrocaudal aspect of the bladder. Both mares survived after treatment for uraemia. Bladder rupture, although uncommon, may affect peripartum mares. Approach to the ruptured bladder without urethral sphincterotomy in a standing position should be considered as a choice for surgical repair.     

Supplemental l-Arginine Shortens Gestation Length and Increases Mare Uterine Blood Flow before and after Parturition

Supplementing diets with l-Arginine (Arg) improves female reproductive performance and reproductive blood flow in other species. The objectives of this study were to investigate uterine artery blood flow changes before and after parturition, and evaluate blood flow in Arg supplemented and control mares by Doppler ultrasonography. Sixteen light-horse mares began Doppler ultrasonography evaluation, 21 days before expected foaling date (EFD) and continued until day 7 postparturition. The mares under treatment (n = 8) were supplemented with 100 g Arg, once daily, beginning with 21 days before EFD. Blood flow measurements were calculated as pulsatility index (PI) and resistance index (RI) for both uterine arteries, either ipsilateral or contralateral; to uterine horn of established pregnancy; defined gravid uterine artery (GUA) and non-gravid uterine artery (NGUA), respectively. The mares under treatment had a shorter gestation length (337 ± 1.7 days) as compared to control (345 ± 2.1 days; P ≤ .05). No differences in gestation length were observed between groups when examined by age, parity, EFD, or sex of foal. Both GUA and NGUA uterine artery diameter decreased from the day before parturition to day 7 after parturition (P ≤ .001). During this time period, both PI and RI increased (P ≤ .01); indicating less blood flow. A treatment effect was observed with Arginine-treated mares having greater blood flow prepartum in the NGUA (P ≤ .001) and postpartum in the GUA (P ≤ .05), for both indices. The data demonstrated that supplementing mares with Arg shortened gestation length and increased uterine arterial blood flow before and after parturition.     

Use of an ecraseur for ovariohysterectomy in mares.

Ovariohysterectomy was performed in 20 mares at three stages of estrus. An ecraseur was used to severe the ovarian branch of the ovarian artery and vein and the ovarian suspensory ligament en masse. All other vessels supplying the ovaries and uterus were doubly ligated and transected. All mares survived. Complications were intraoperative hemorrhage in three mares, postoperative vaginal bleeding in two mares, and a hematoma in the remnant of the broad ligament in one mare. No adhesions between the uterine stump or remnants of the broad ligament and abdominal structures were detected by palpation per rectum.     

Infertility in a mare with a chronic uterine tear,diagnosis and successful standing hand‐assisted laparoscopic repair

Uterine rupture is a well recognised and potentially fatal complication in the peripartum broodmare. This case report describes a 16 month history of infertility as the sole clinical sign in a Thoroughbred mare, subsequently diagnosed with a full thickness uterine tear. The clinical course, standing laparoscopically assisted repair and outcome are discussed.     

Impotence in the bull: (3) Rupture of the corpus cavernosum penis proximal to the sigmoid flexure

Penile haematomas, lying outside the tunica albuginea, were found in six bulls of various breeds, aged one-and-a-half to nine-and-three-quarter years. In five cases the haematomas definitely arose from rupture of the dorsal or crural canal and tunica albuginea of the corpus cavernosum penis (ccp) within the proximal 12 cm of the penile body. In the sixth bull, lesions of the tunica albuginea and distortion of the penis just proximal to the proximal bend of the sigmoid flexure may have represented the site of rupture. There was no evidence of rupture at the distal bend of the sigmoid flexure or of rupture of the dorsal penile vessels in any of these specimens. In all six specimens, the dorsal canals of the ccp were occluded by translucent fibrous tissue distal to the sites of rupture; this was considered to be the factor immediately predisposing to proximal rupture of the ccp. Clinically, a small haematoma in the perineal region is difficult or impossible to palpate externally. However, when the haematoma of proximal origin is large, differentiation from rupture at the distal bend of the sigmoid flexure may be based tentatively on a careful clinical examination. In cases of proximal rupture the haematoma lies caudal and dorsal to the scrotum and no abnormality of the distal bend may be palpable. In cases of distal rupture, the haematoma usually lies cranial to the scrotum and involves the distal bend of the flexure. Severe disruption of the vascular structure is found in cases of proximal rupture of the ccp; therefore the prognosis is grave.     

A review of mare endometritis in Iran

One hundred eleven mares were examined and selected on the basis of some criteria (history of recent genital discharge and/or abortion, dystocia, retained placenta, rectovaginal rupture and/or failing to conceive after repeated services), rectal palpation, and ultrasound examination of genital tract. The mares were classified in 3 groups (young, middle age and old). During estrus, a specimen was taken by uterine swab and endometrial biopsy. Endometritis was diagnosed and graded on the basis of pathology results and treatment was done on the basis of culture and antibiogram tests. At the first step of treatment, uterine lavage with warm saline followed by intrauterine administration of a specific antibiotic, or povidone iodine solution, was done daily during estrus. If the mare had not responded to the first treatment, in the next steps systemic antibiotic therapy was also added to the mentioned treatment. The results showed that Escherichia coli was the most frequently isolated pathogen and response to the treatments was significantly better in the young mares with category IIA fibrosis of uterine pathology than that of two other groups. It was also found that there is no relationship among age and kind of pathogen to mare endometritis.

Introduction

Bacterial endometritis is the most common cause of subfertility.[1, 2, 3 and 4] In 1992, Kenney argued against the general use of the term “endometritis” to describe the two often quite separate conditions of endometrium: the degenerative changes often associated with age and parity. He suggested the use of the term “endometriosis” instead of the term “chronic degenerative endometritis” (CDE): the inflammatory infection that is divided to “acute endometritis” and “chronic infiltrative endometritis” (CIE). [5]In some mares, the bacteria becomes established and uterine infection develops in which the sources of uterine contamination include coitus, parturition, and reproductive examination.[2, 4 and 6] These mares remain persistently infected and are termed “susceptible” mares, which may have conformation defects such as pneumovagina, uterine pooling, and foaling injuries, which predispose them to endometritis. [7] There are some data indicating that age, parity, and barren years may have important bearing on the breeding prognosis of the mare. [8 and 9] In general, loss of resistance to infection is associated with advancing age and multiparity, factors that are frequently associated with increasing value of mares. [10] By far, Streptococcus zooepidemicus followed by E coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae are the most frequently isolated pathogens,[7 and 10] but there are a few reports that E coli, K pneumoniae, and Corynebacterium spp. are the most frequent isolates.[4 and 11] Endometrial cytology and culture samples often reveal evidence of inflammation and infection. To improve reproductive management of mares in Iran, this study was conducted to assess main causes of mare endometritis and also the relation of age and kind of pathogen to mare endometritis, plus the efficacy of treatment methods in problem mares.

Materials and methods

One hundred eleven mares of different breeds (Thoroughbred, Arab, cross-bred, and two native breeds including Kord and Turkaman) from 4 to 26 years of age were examined and selected during April 1996 to June 2001. They had a history of recent genital discharge and/or abortion, dystocia, retained placenta, rectovaginal rupture, and/or failing to conceive after repeated services. The sanitary condition of the foaling boxes was also noted. The mares were classified in three groups according to the age: (1) young (4-9 years old), (2) middle age (10-14 years old), and (3) old age (≥15 years old).Each mare was restrained in a stock. The vulvar discharge and pneuomovagina were recorded if present. Rectal palpation and ultrasound examination of genital tract were done to determine uterine tone and presence of uterine fluid. During the estrus, a double-guarded swab was passed per vagina into the uterus. The swab was kept in contact with the endometrium for a minimum of 20 seconds and was placed in transport media. In the laboratory the swab was cultured as described by Rickets.[12] An endometrial biopsy specimen was also taken, fixed, and processed as described by Rickets. [13] Endometritis was graded as described by Kenny and Doig. [14] Briefly, they are Grade I (Absent), Grade IIA (Mild), Grade IIB (Moderate), and Grade III (Severe). After specifying the causal organism of endometritis, treatment was done on the basis of culture and antibiogram tests as described by Asbury et al, [10] the antibiotics used for treatment were: gentamicin (intrauterine administration, 2-3 g), amikacin (2 g, intrauterine administration), potassium penicillin G (5 million units, intrauterine administration), and chloramphenicol (3 g, intrauterine administration). [10 and 15] A diluted solution of povidone iodine (0.2%) as uterine lavage was also used in cases whose uterine culture results were yeasts. The following protocol was carried out to treat bacterial endometritis at the first step:Uterine lavage with warm saline followed by intrauterine administration of specific antibiotic or povidone iodine (on the basis of uterine culture and antibiotic sensitivity test results) was done daily during estrus. Casslick operation was done in pneuomovaginal cases. In the next estrus, the uteri were examined by rectal palpation and ultrasonography procedure. If they were involuted, had enough tone and had no fluid, the mares were clinically defined to be treated and bred artificially with fresh semen collected from an approved fertile stallion (about 500 million progressive motile sperm) in Kenney's extender. If there was recurrent endometritis or the uterus was not involuted and/or had not enough tone or had some fluid in it, another specimen was taken by a double-guarded swab. On the basis of culture and antibiotic sensitivity test results, mares that had not responded to the first treatment were treated again with the first step protocol plus systemic antibiotic therapy (gentamicin, amikacin, or procaine penicillin G in bacterial-caused endometritis) or just flushed the uterus with saline and 0.2% povidone iodine solution (in yeast-caused endometritis and/or endometriosis). This treatment was also carried out daily during the estrus. Some of the mares that had not responded to the second treatment protocol were treated again (based on uterine culture and antibiotic sensitivity test results) for one to two times more in the next estrus phases as in the second step. The data were analyzed by χ² test.

Results

Table 1 shows the pathogens isolated in three successive cultures from the uteri of the mares. Among 197 swabs were taken from the uteri of 111 mares, the cultures were positive in 175 instances in which the mixed organisms were isolated in 20 cases. The interesting point in these 3 successive cultures was the persistence of one Klebsiella infection detected in a mare belonging to the second group, in the all-performed treatments. The isolated P aeruginosa in a mare belonging to the third group had the same result. The combination of Candia albicans with E coli and/or E coli with Enterobacteriaceae were the most common feature of mixed micro-organisms isolated in the culture plates.     

Use of a Mycobacterial Cell Wall Extract (MCWE) in Susceptible Mares to Clear Experimentally Induced Endometritis With Streptococcus zooepidemicus

The ability of an immunomodulator, mycobacterial cell wall extract (MCWE), to clear uterine infection in susceptible mares after an experimental challenge withStreptococcus zooepidemicus was evaluated. Thirty mares susceptible to endometritis, based on the presence of uterine fluid during both diestrus and estrus, were selected from a herd of 896 and inoculated with a live culture of 5 × 10⁶ CFU of S. zooepidemicus on day 1 of estrus. Twenty-four hours later, mares were evaluated by ultrasonography, bacteriology, exfoliative cytology, and uterine biopsy to confirm infection. Forty-eight hours after inoculation, and on confirmation of uterine infection, mares were randomly assigned to one of four unbalanced experimental treatments to receive 1500 μg MCWE IU (n = 10) or IV (n = 10), or placebo IU (n = 5) or IV (n = 5). Mares were examined at ovulation and 7 days post-ovulation for uterine fluid via transrectal ultrasonography and for bacteriology, exfoliative cytology, and uterine biopsy. Efficacy was based on the ability of the mare to clear endometritis as determined by negative bacteriology and reduced numbers of polymorphonuclear cells (PMNs) on uterine biopsy. Because no statistical difference was detected between routes of administration on day 7 post-ovulation, the data sets were combined and re-analyzed to evaluate overall efficacy. Endometritis was observed in all placebo-treated mares 7 days post-ovulation, whereas treatment with MCWE resulted in the elimination of endometritis in 35% of the mares by the time of ovulation, and 70% of the mares by 7 days post-ovulation. Treatment with MCWE, compared with the placebo group, resulted in a significant decrease in the number of mares positive for endometritis at ovulation based on exfoliative cytology and bacteriology (P < .01) and at 7 days post-ovulation based on biopsy, exfoliative cytology, and bacteriology (P < .001). Results indicate that MCWE was an effective treatment for the elimination of endometritis caused by S. zooepidemicus in mares.     

Actions of isoflurane and halothane in pregnant mares.

Eighteen healthy, pregnant mares scheduled for laparotomy and uterine manipulation were randomly allotted to 2 equal groups. After IV administration of xylazine hydrochloride and thiamylal sodium, general anesthesia was maintained with halothane (HALO) or isoflurane (ISO) in oxygen. Results of cardiovascular measurements were similar with both inhalant anesthetics; mean arterial blood pressure was 79 and 82 mm of Hg with HALO and ISO, respectively. Respiratory rate decreased most with ISO (mean frequency was 4 and 9 breaths/min with ISO and HALO, respectively). Partial pressure of arterial CO2 was increased similarly with HALO and ISO. Partial pressure of arterial O2 varied greatly among mares and decreased with duration of use of both anesthetics. Recovery time from anesthesia was significantly (P < 0.05) shorter after use of ISO vs HALO. Minor superficial injuries were associated with recovery from both anesthetics (in 5 mares with ISO and in 1 mare with HALO). Physical signs of postanesthetic myopathy or vital-organ dysfunction were not associated with either agent.     

Total and partial ovariohysterectomy in seven mares

REASONS FOR PERFORMING STUDY: Ovariohysterectomy appears to have a low mortality rate in mares, but the procedure needs to be reviewed because of the high risk of life-threatening complications. HYPOTHESIS: That ovariohysterectomy can be effective treatment for a variety of uterine diseases in mares and carries a good prognosis. METHODS: Diagnosis, clinical data, surgical technique, post operative care, complications and outcome were recorded from medical records of 7 mares that underwent total (6) and partial (1) ovariohysterectomy at the University of Illinois from 1994 to 2001. RESULTS: The indications for ovariohysterectomy were chronic pyometra (4 mares), chronic uterine torsion (n = 2) and chronic intramural haematoma (n = 1). Surgical exposure was difficult but was improved by traction on stay sutures and right-angled clamps. In some cases, application of the TA-90 autosuture instrument as a right-angled clamp to the caudal part of the uterus improved access to the uterine stump. The most common post operative complications were decreased faecal output, decreased intestinal sounds (4 mares) and mild abdominal pain (2). Two mares had mild to moderate incisional infections. Other previously reported complications, such as haemorrhage, septic peritonitis, uterine stump infection or necrosis, and diarrhoea, did not occur. All mares survived over follow-up periods of 6 months to 5 years and were used for riding (6 mares) and embryo transfer (1 mare, after partial ovariohysterectomy). CONCLUSIONS AND POTENTIAL RELEVANCE: According to this study, the prognosis for mares after ovariohysterectomy appears to be good, despite the technical difficulties of the procedure. The prevalence of life-threatening complications can be lower than reported.     

The effects of an advanced uterine environment on embryonic survival in the mare

Reasons for performing the study: During embryo transfer (ET) the equine embryo can tolerate a wide degree of negative asynchrony but positive asynchrony of >2 days usually results in embryonic death. There is still confusion over whether this is due to the inability of the embryo to induce luteostasis or to an inappropriate uterine environment. Objectives: To assess embryo survival and development in an advanced uterine environment. Hypothesis: Embryo–uterine asynchrony, not the embryo's inability to induce luteostasis, is responsible for embryonic death in recipient mares with a >2 days chronologically advanced uterus. Methods: Experiment 1: Thirteen Day 7 embryos were transferred to the uteri of recipient mares with luteal prolongation, occasioned by manual crushing of their own conceptus, such that donor–recipient asynchrony was between +13 and +49 days. Experiment 2: Day 7 embryos were transferred to recipient mares carrying their own conceptus at Days 18 (n = 2), 15 (n = 2), 14 (n = 4), 12 (n = 4) or 11 (n = 4) of gestation. In addition, Day 8 embryos were transferred to 4 pregnant recipient mares on Day 11 of gestation. Results: No pregnancies resulted following transfer of Day 7 embryos to recipients in prolonged dioestrus with asynchronies between +13 and +49 days. However, the use of early pregnant mares as recipients resulted in 5/20 (25%) twin pregnancies, 4 of which came from the transfer of a Day 8 embryo to a Day 11 recipient. All transferred embryos showed retarded growth, with death occurring in 4/5 (80%). Conclusions and potential relevance: The results emphasise the importance of an appropriate uterine environment for embryo growth and the inability of equine embryos to survive transfer to a uterus >2 days advanced even when luteostasis is achieved. It is possible that in normal, non‐ET equine pregnancy, embryo–uterine asynchrony may account for some cases of embryonic death.     

Post-partum uterine rupture: Standing repair in three mares using a laparoscopic technique

Three mares underwent diagnostic laparoscopy because of suspicion of post-partum uterine ruptures. All three horses showed clinical signs of a uterine rupture between 1 and 3 days after parturition and underwent diagnostic laparoscopy. In all cases a full thickness uterine rupture could be detected and was sutured laparoscopically. Availability of suture material and surgeon experience were responsible for the surgical methods chosen for repair. In the first case, a hand-assisted laparoscopic approach was chosen for suturing the ruptured uterus, whereas in the other cases the approach was entirely laparoscopic. In the second case, extracorporeal knots were used for the repair and in the last case described a barbed loop suture was available for closure of the uterus. Two of three mares were alive for at least 12 months after surgery without any abdominal problems. One of these mares delivered a healthy foal 2 years after surgery. The remaining mare died 3 months after surgery but no necropsy was done. Laparoscopy should be considered for post-partum mares with signs of peritonitis to access the uterus and repair a rupture if it is accessible. A laparoscopic approach using intracorporeal knots or barbed sutures for the repair of the uterine rupture as well as a hand-assisted laparoscopic approach are feasible. The use of the barbed suture for intracorporeal closure makes the minimal invasive laparoscopic technique easier to perform.     

Caudal proximal tibial deformity and cranial cruciate ligament rupture in small-breed dogs

Eight dogs presented with chronic hindlimb lameness associated with cranial cruciate ligament rupture. Seven were small terriers. A caudal deformity of the proximal tibial shaft, originating at the proximal tibial physis, and an excessive caudal slope of the tibial plateau were present bilaterally in all dogs. The deformity was thought to be responsible for the cranial cruciate ligament failure and poor response to conservative management. Tibial plateau angles were in excess of 26 degrees in all dogs. The lameness was bilateral in three dogs. There was complete cranial cruciate ligament rupture in seven stifles and partial rupture in four. There were no meniscal injuries. Surgical correction resulted in a significant improvement (P<0.0001) in all dogs, with a mean follow-up of 12 months (range three to 24 months). There were no complications.     

Qualitative analysis and functional classification of the uterine proteome of mares in oestrus and dioestrus

Quantitative analysis of the uterine flush fluid proteome of mares in oestrus and dioestrus has been previously reported. The objectives of this study were to: a) evaluate qualitative differences in the uterine flush fluid proteome between mares in oestrus and mares in dioestrus and b) perform a functional classification of proteins either unique to each stage or common between the two stages. Uterine flush fluid samples were collected from 8 light breed mares in either oestrus (n = 5) or dioestrus (n = 3). Proteomic analysis of the samples was conducted using liquid chromatography–tandem mass spectrometry. Proteins exclusively detected in oestrus or dioestrus and those common to both stages were identified using the Scaffold software (version 4.4.8, Proteome Software Inc., Portland, OR). The identified proteins were classified into gene ontology (GO) categories (cellular component [CC], molecular function [MF] and biological process [BP]) using the PANTHER ( www.pantherdb.org ) classification system version 14.0. Of 172 proteins identified, 51 and 28 were exclusively detected in mares in oestrus and dioestrus, respectively, and 93 proteins were common to both stages. The most represented terms in various GO categories were similar among the three subsets of proteins. The most represented CC terms were extracellular region and cell, the most represented MF terms were catalytic activity and binding, and the most represented BP terms were metabolic process and cellular process. In conclusion, proteomic analysis of the uterine flush fluid enabled the identification of subsets of proteins unique to oestrus or dioestrus, or common to both stages. The results of this study can serve as a baseline for future research focused on finding stage-specific protein markers or evaluating differences in the uterine flush fluid proteome between normal mares and those with uterine disease.     

Spectral Doppler ultrasound in selecting an equine embryo receiver

The aim of this study was to evaluate whether the RI and PI values would help in choosing the best embryo recipient, and observe whether CL vascularization would influence P4 production. During the breeding season 2018/2019, the study was conducted using 35 mares, which is used for reference to collect data for the project on the day of embryo transfer. The utilized mares were divided into five groups followed by the day after ovulation, with D0 being the day of ovulation. Therefore, the five groups are as follows: D4—mares that were on the 4th post-ovulation day; D5—mares that were on the 5th post-ovulation day; and doing so successively for D6, D7 and D8. On the day of embryo transfer, the CL of the mares that selected as recipients was evaluated by B-mode and power flow mode ultrasonography and the right and left dorsal branches of the uterine arteries by spectral Doppler ultrasonography. Blood samples were taken on the day of the embryo transfer for a dosage of P4 concentration by radioimmunoassay. No statistical difference was found between the variables when the mares were separated into pregnant and non-pregnant mares, or when they were separated by age groups. When the groups of mares were compared by the day of embryo transfer, the statistical difference was found between the groups D5 × D6 (p = .0053) and D6 × D8 (p = .0036) in RI variable. In PI variable, the statistical difference was found between the groups D4 × D8 (p = .049), D5 × D6 (p = .0446) and D6 × D8 (p = .0024). We conclude that the mares with RI measurement of uterine arteries near 1.0 are correlated to mares with high CL vascularization and elevated P4 concentration.     

Proteins and Enzymes in Uterine Lavage Fluid of Postpartum and Nonparturient Mares

Uterine lavage fluids from postpartum and nonparturient mares were compared to determine when the normal secretory capacity of the postpartum uterus is restored. Lavage fluids were obtained from cyclic nonparturient mares on the second, fourth or fifth day of oestrus, and 3, 8, or 14 days after ovulation (seven mares/sampling day). Twelve intact postpartum mares were sampled 1 to 28 days postpartum (group A: 1, 6, 12 and 20; group B: 2, 8, 14 and 24; group C: 4, 10, 16 and 28 days postpartum; four mares/group). Three ovariectomized (OVX) postpartum mares were sampled as mares in group C. Samples were analysed for neutrophils, bacteria, total protein concentration, proteolytic and antiproteolytic activities and for various lysosomal enzyme activities. In nonparturient mares, activities of acid phosphatase, β‐glucuronidase (B‐Gase), and N‐acetyl‐β‐D‐glucosaminidase (NAGase) in uterine lavage fluids were significantly higher in mid‐ and late‐dioestrus than in mid‐ to late‐oestrus (p < 0.05). Lysozyme concentration, trypsin‐inhibitor capacity (TIC), and plasmin activity were below the detection limit in nonparturient mares. One to four days postpartum, total protein, acid phosphatase, B‐Gase, and NAGase were high but declined rapidly thereafter. Lysozyme and plasmin activities were high 1 to 6 days postpartum. TIC peaked around day 6 postpartum. On day 16 postpartum, acid phosphatase, B‐Gase, and NAGase, being progesterone‐dependent, tended to be higher in intact mares than in OVX ones (p < 0.1). Total protein and lysozyme concentrations, TIC, and B‐Gase (p < 0.01) and acid phosphatase (p < 0.05) activities were significantly higher in parturient mares during postpartum oestrus than in oestrous nonparturient mares. High total protein concentration and TIC, and detectable lysozyme and plasmin activities during postpartum oestrus were associated with uterine inflammation. During dioestrus, differences between postpartum and nonparturient mares were not statistically significant and suggested that the endometrium of postpartum mares had resumed its normal secretory capacity by this time.     

Using “Uterine Index” to diagnose embryonic death in mares

Embryonic death plays an important role in infertility in mares, and most of embryonic mortality occurs at an early stage (before day 40) of pregnancy. Previous investigations focused on B-mode sonographic images to determine abnormal pregnancy; however, no study has provided a quantitative and objective method to diagnose embryonic death. Here we developed a new index, “Uterine Index,” using color Doppler sonography to describe alterations of uterine blood flow in physiological and pathological gestations during early gestation. Six Trotter mares were used in this study. The pregnancies were interrupted with a prostaglandin F2α (PGF_2α) analog (tiaprost) at days 25, 30, and 40. Three gestations were investigated in each stage. After the PGF_2α administration, these mares were examined every 8 hours until the embryonic vesicles were not visible on ultrasound. In the control group, three pregnancies were examined daily from days 12 to 60. The embryo size was measured in B-mode sonography, and the blood flow in uterine arteries was examined in Doppler ultrasound. The resistance index (RI) and uterine index (UI) were calculated. In the control group, the RI values of uterine artery were lower in pregnant side compared with that of the non-pregnant side from day 15 on, and the difference became greater as the pregnancy advanced (P < .05). The UI was close to 0 before week 3 and increased to approximately 10 after week 4. In the PGF_2α-treated group, the UI values reduced from pregnancy level to approximately 0. As early as 32 hours before embryonic death, we observed a significant difference of UI values between control and treated mares. On the contrary, the embryo size of treated mares was not different from that of normal mares. Through the analysis of all recorded UI values, we suggest that the UI values 10 and 5 are good values for distinguishing normal pregnancy (UI >10) from embryonic death (UI < 5). If the UI values are located between 10 and 5, a reexamination at intervals of 1 to 3 days is recommended. Our findings demonstrate that the UI is a useful method to diagnose and predict embryonic death in mares by providing reliable and objective information.     

Common Peripartum Problems in the Mare

Reproductive problems in the perinatal period can adversely affect a mare's future fertility or, worse, may be life threatening to her or her foal. A number of abnormalities can occur, including uterine torsion, uterine artery or uterine hemorrhage, retained fetal membranes, metritis, rectovaginal injuries, and necrotic vaginitis. Because hemorrhage, retained placenta, metritis, and necrotic vaginitis can happen after a normal delivery or after dystocia, all mares should be monitored closely in the first days after parturition for signs of lethargy, depression, colic, sore feet, or anorexia. Methods for diagnosing, managing, and treating these conditions and the possible complications are discussed.     

RADIOGRAPHIC AND ULTRASONOGRAPHIC EVALUATION OF THE PATELLAR LIGAMENT FOLLOWING TIBIAL TUBEROSITY ADVANCEMENT

Effect of tibial tuberosity advancement (TTA) on the patellar ligament has not been described. Our purpose was to evaluate the patellar ligament radiographically and ultrasonographically before and after a TTA. Twenty‐one stifles (20 dogs) were evaluated preoperatively (T0), and at six (n=18) (T1) and 16 weeks (n=17) (T2) postTTA. Radiographically, proximal and distal thickness of the patellar ligament was assessed and a ratio to the total length of the ligament was calculated to compensate for the magnification. Ultrasound evaluation included measurements of the transverse thickness and cross‐sectional area at three different levels, as well as a subjective score of ligament changes. In comparison with T0, all radiographic and ultrasonographic measurements increased significantly, 6 weeks postoperatively (P≤0.04), and did not change 16 weeks postoperatively compared with T1. The subjective score worsened significantly from T0 to T1 and T0 to T2 (P<0.0001), and improved significantly from T1 to T2 (P=0.02). Larger cage size was associated with a more severe increase in radiographic proximal thickness to total length ratio and ultrasonographic middle transverse area at both follow‐up examinations (P0.02). Dogs in which arthrotomy was not performed appeared to have ultrasonographically less changes. In conclusion, patellar desmopathy was a common postoperative sequel to TTA. Surgical trauma, arthrotomy, perfusion injury, complete vs. partial cranial cruciate ligament rupture, larger tibial advancement, postoperative activity or altered insertion angle of the patellar ligament at the tibial tuberosity are suggested causes, that should be elucidated in a larger study cohort.     

Retrospective study on the incidence of postinsemination uterine fluid in mares inseminated with frozen/thawed semen

Uterine fluid accumulation has been reported after insemination or natural breeding of mares. This retrospective study examined the factors affecting the incidence of uterine fluid after insemination of frozen semen. Specifically, this study determined the association between mare age, reproductive status, fluid accumulation, and pregnancy rates in mares. Records were available from 283 warmblood mares throughout 496 cycles. Mares were divided into maiden, foaling, and barren and age groups of 3 to 9, 10 to 16, and more than 16 years. Mares were inseminated only once with frozen semen within 4 to 8 hours before or after ovulation. Ultrasound examinations were performed 12 to 18 hours after insemination. A depth of at least 20 mm of fluid was considered significant. Mares with less than 20 mm were treated with oxytocin, and those with more than 20mm of fluid were given oxytocin and uterine lavage. Pregnancy determination was performed at 14 to 16 and 30 to 50 days after ovulation. Fluid level of more than 20 mm was recorded in 25% of the cycles. Barren mares and aged mares (10-16 and > 16 years) had a higher incidence of uterine fluid accumulations. Per-cycle pregnancy rate was lower (45%) in mares with uterine fluid than in mares without uterine fluid (51%). This difference was primarily due to the reduction in fertility of mares who were older than 16 years and retained fluid after insemination. Apparently, oxytocin and lavage treatments provided acceptable fertility in the other groups of mares that had uterine fluid.

Introduction

Use of equine frozen semen is accepted by the majority of horse registries. According to several field studies,[1, 2, 3, 4 and 5] insemination of frozen semen has resulted in acceptable pregnancy rates. Postbreeding fluid accumulation is a physiologic inflammation that clears the uterus of foreign material such as excess spermatozoa, seminal plasma, bacteria, and extenders. [6, 7, 8, 9 and 10] Uterine fluid can be easily diagnosed with ultrasonography. [10, 11 and 12] Persistent postbreeding uterine fluid has been associated with a decrease in fertility after natural mating or artificial insemination (AI) of fresh semen. [11, 12 and 13] Predisposing factors to persistent fluid accumulations are reduced myometrial contractions, poor lymphatic drainage, large overstretched uterus, and cervical incompetence. [7, 14 and 15] Normal mares are able to expel uterine fluid quickly after inseminations, whereas susceptible mares accumulate fluid in their uterine lumen for more than 12 hours after breeding or insemination. [10]It is commonly stated that insemination with frozen semen leads to greater post-AI fluid accumulation than insemination with fresh or cooled semen or after natural mating. Apparently, there is only 1 controlled study on this comparison.[7] The authors reported that infusion of frozen semen resulted in a greater inflammatory response than natural breeding. In a field study, [16] 16% of mares naturally mated had persistent postbreeding fluid accumulations compared with a 30% rate reported for mares inseminated with frozen semen. [1 and 2] More recently, Watson et al. [17] reported a postbreeding fluid accumulation rate of 16%, which is identical to that reported for natural mating. [16] It is difficult to compare studies because details of mare selection and insemination or breeding frequencies are not always reported. Obviously, a higher proportion of barren and aged mares in a study would increase the incidence of postbreeding fluid accumulation. [1 and 2]The study presented herein was a retrospective study designed to determine the incidence of postbreeding fluid accumulation in a large number of mares inseminated with frozen semen. Associations were determined between mare age, reproductive status and fluid accumulation, and pregnancy rate in mares with and without uterine fluid accumulation.

Materials and methods

Mares

Records were available from 283 warmblood mares inseminated with frozen semen at the Cristella Veterinary Clinic in Italy during 1998 to 2001. Mares ranging in age from 3 to 20 years were inseminated with semen that was frozen in 10 centers and was from 34 stallions. The broodmare population was subdivided into 3 reproductive groups: 89 maiden mares (mean age, 7.2 years), 106 foaling mares (mean age, 9.4 years), and 87 barren mares (mean age, 11.9 years). Maiden mares older than 7 years were selected with biopsy scores of 1 or 2 only. Barren mares were open for no more than 2 consecutive seasons and had negative cytology and bacteriology scores. Age groups were divided as follows: 3 to 9 years (n = 132), 10 to 16 years (n = 137) and older than 16 years (n = 14). Data from 496 cycles were used. Distribution of the estrous cycles was 172, 157, and 167 in the maiden, foaling, and barren groups, respectively; and 224, 244, and 28 in the youngest, intermediate, and oldest groups, respectively.

Mare reproductive management and artificial insemination protocol

During estrus, all mares underwent a daily ultrasound examination with a 5-mHz transrectal probe (SA 600 Vet; Medison Inc., Seoul, South Korea) until 1 or more 35-mm ovarian follicles were detected. Ovulation was then induced by the intravenous administration of 2000 IU of human chorionic gonadotropin (hCG). Ultrasound examination was performed 12 hours after hCG treatment and then every 4 to 8 hours until ovulation occurred. Mares were inseminated only once within a period of 4 to 8 hours before or after ovulation. The semen used was thawed according to the distribution center's instructions and had the following minimum post-thaw quality requirements: not less than 200 × 10⁶ progressively motile spermatozoa per dose and a minimum of 30% progressive spermatozoal motility. Foaling mares were not inseminated at their first postpartum (“foal heat”) estrous period, because pregnancy rates are recognized to be lower than during the subsequent estrous periods.[18] During the first postpartum estrus, ovarian ultrasound scan examinations were performed every 2 to 3 days until an ovulation was detected. A prostaglandin F₂α injection was given 5 days later to short-cycle the mare.

Postinsemination monitoring

An ultrasound examination of the reproductive tract was performed 12 to 18 hours after insemination to detect any intrauterine fluid accumulation. The presence and depth of intrauterine fluid was recorded. Twenty millimeters or more of grade II or III intrauterine fluid[19] was recorded as a significant amount of fluid. Mares with less than 20 mm of fluid were treated with an intravenous injection of 20 IU oxytocin. For mares with more than 20 mm of fluid, oxytocin was administered, and the uterus was flushed daily with buffered saline solution: 1-L aliquots were infused and recovered until the recovered fluid was clear. In these mares, oxytocin treatment was repeated up to 3 times daily. Post insemination treatments were performed for no more than 4 days after ovulation had occurred.Pregnancy diagnosis was performed with ultrasound at 14 to 16 days after ovulation. Scans were then repeated at 30 and 50 days of gestation to confirm the presence in the uterus of an apparently healthy developing conceptus.

Statistical analysis

χ² Analysis was used to determine the effect of reproductive status and age on the incidence of fluid accumulation. In addition, the influence of persistent uterine fluid accumulation on pregnancy rates per cycle was determined for each reproductive class and age by using χ² analysis.

Results

The per-cycle pregnancy rate at 14-16 days after ovulation was 49.3% (245/496 cycles). By the end of the season, 245 of 283 mares (86.5%) were confirmed pregnant. Fluid level of at least 20 mm (grade II or III) was recorded in 126 of the 496 cycles (25.4%). Barren mares had a higher (P < .05) incidence of postbreeding fluid accumulation (64/167; 38.3%) than maiden (34/172; 19.7%) and foaling (28/157, 17.8%; Table 1) mares. The incidence of fluid accumulation was also higher in mares older than 16 years (19/28; 67.8%) than those aged 10 to 16 years (69/244; 28.2%) and 3 to 9 years (38/224; 17%). The incidence of uterine fluid was also higher (P < .05) for mares aged 10 to 16 years than those aged 3 to 9 years (Table 2). Overall, the per-cycle pregnancy rate was lower (P < .05) for mares with post-AI fluid accumulations than for those with no uterine fluid or only a small quantity of fluid (57/126, 41.9% vs 188/360, 56.2%). Pregnancy rates were similar (P > .05) for mares with or without uterine fluid when comparisons were made within maiden and barren mare groups. However, more foaling mares became pregnant when no fluid was detected after insemination. Pregnancy rate for this group (68.1%) was higher than that for maiden (44.2%) and barren (44.6%) mares (Table 3). Older mares with uterine fluid accumulations had a lower per-cycle pregnancy rate (36.8%) than mares in the same group but without fluid. Surprisingly, if no fluid was detected, the highest pregnancy rates were in mares older than 16 years ( Table 4).