Consequences of Intrauterine Enrofloxacin Infusion on Mare Endometrium

Endometritis is an important cause of infertility in mares. Enrofloxacin is a broad-spectrum antibiotic to which most equine endometritis pathogens are not resistant. The objective of this study was to determine whether enrofloxacin is safe to use as a conventional intrauterine infusion treatment. Nine healthy mares received intrauterine infusions of enrofloxacin (Baytril 100, 100 mg/mL, Bayer Health Care LLC, Animal Health Division) at 2.5 mg/kg daily for 3 days. Ultrasonographic examination and vaginal examinations were performed during the study. Endometrial biopsies were performed before treatment (S0) and 24 hours after the last treatment (S1) to evaluate acute effects. For evaluating chronic effects, biopsies were performed at 14 days (S2) and 60 days posttreatment (S3). Biopsies were graded histologically by the Kenny and Doig category scale. Difference in histological biopsy grade before and after treatment was compared between biopsies by using a repeated-measures one-way analysis of variance. and significant changes in grades were used to assess treatment effects. The vaginal and ultrasonographic examination after intrauterine infusion of enrofloxacin showed that all mares had severe purulent vaginitis and uterine fluid accumulation of ≥2 cm, with ≥1.5-cm thickening of the endometrial wall which persisted in most mares until the end of the study. Histologically, there was acute endometrial ulceration, necrosis, and hemorrhage in biopsy S1 in all mares, categorized as grade III. In biopsy S2, most mares developed fibrosis and inflammation graded as IIb (four of nine mares) or III (four of nine mares). In biopsy S3, fibrosis was extensive and had variable inflammation, graded as IIb (two of nine mares) or III (five of nine mares), with some mares healing to grade IIa (two of nine mares). There was an overall worsening of endometrial biopsy grade from I to III at S3 compared with S0 (P < .001). These results confirm that enrofloxacin is not suitable for conventional intrauterine infusion treatment in mares.     

Technique and interpretation of cervical and endometrial cytology in the mare

This paper illustrates and describes the findings obtained from cervical and uterine cytology smears in the mare. Cervical/endometrial cytology is a simple technique that provides valuable information to the clinician regarding the existence of an acute or active endometritis. The most frequent observation is the presence of neutrophils. Chronically infected/inflamed mares with some degree of inflammatory cellular infiltration can also be identified on the cytology smear by the presence of neutrophils, macrophages and lymphocytes. Microorganisms are sometimes found with either acute or chronic endometritis.

This technique is not a substitute for uterine culture and biopsy. The correct identification of bacteria should be based on the cultural characteristics of the microorganisms.     

Effect of a povidone-iodine intrauterine infusion on progesterone levels and endometrial steroid receptor expression in mares

Background

Intrauterine infusions have been widely used for the treatment of endometritis in the mare. Nevertheless, their consequences on endocrine and endometrial molecular aspects are unknown. We studied the effect of a 1% povidone-iodine solution intrauterine infusion on progesterone levels, endometrial histology and estrogen (ERα) and progesterone (PR) receptor distribution by immunohistochemistry.

Methods

Fourteen healthy mares were used in this study. Estruses were synchronized and seven mares were treated with intrauterine infusions at days 0 and 2 post ovulation of two consecutive estrous cycles. Uterine biopsy samples were taken on days 6 and 15 post ovulation.

Results

The treatment did not induce an inflammatory response indicating endometritis, neither affected the ERα. However, it reduced the percentage of PR positive cells (PPC) on day 6 (deep glandular epithelium, control: 95.7 vs. infused: 61.5, P < 0.05). Treated mares tended to have lower progesterone levels on day 2 (3.9 ng/ml vs. 6.6 ng/ml, P = 0.07), and higher levels on day 15 compared with controls (4.4 ng/ml vs. 1.3 ng/ml, P = 0.07).

Conclusion

a 1% povidone-iodine infusion during days 0 and 2 post ovulation in healthy mares did not induce histological changes indicating endometritis, but altered progesterone concentrations and reduced the expression of endometrial PR at day 6 without affecting the ERα. These changes could reduce embryo survival.     

Early foetal death in the mare. Histological, bacteriological and cytological findings in the endometrium.

Fifteen mares which had each lost their pregnancies 2 to 8 times during the preceding 6 years were used in a study comprising 1 or 2 breeding seasons for each mare. During the research period all 15 mares conceived and 10 of the mares had normal pregnancies in the 1st experimental year. Five mares resorbed/aborted once or twice during the 1st year. This was followed by a pregnancy that terminated in a live foal. Histopathological examinations of uterine biopsies showed a wide range of histopathological conditions, from absence of changes excessive for the mare's age and parity to the most severe changes of the endometrium. Most likely, endometritis, acute as well as chronic degenerative endometritis, is the most common cause of early foetal death in the mare. Good breeding management is essential and was often enough to make the mares produce a foal. Additional treatment was needed in some of the mares and was based on the course of the conceptus loss and the condition of the endometrium, as determined by clinical, histological, bacteriological and cytological examinations.     

Occurrence of Intrauterine Purulent Concrements in a Maiden Mare—A Case Report

Pyometra is an uncommon condition in mares associated with various symptoms. Here, we report a case of a 13-year-old Icelandic barren maiden mare with recurrent vaginal discharge. Ultrasonographically, the mare displayed intrauterine spherical masses of inhomogenous texture, which were identified as purulent concrements in hysteroscopy. The purulent concrements were successfully removed via uterine lavage after endoscope-assisted comminution. Microbiologic examination of the concrements revealed growth of Streptococcus equi subspecies zooepidemicus, Actinobacillus species, Pseudomonas aeruginosa, Staphylococcus intermedius, Pseudomonas fulva, Citrobacter freundii, and Chryseobacterium species. Systemic antibiotic treatment with trimethoprim-sulfadiazine and additional uterine lavages were performed for 10 days. A follow-up examination revealed absence of intrauterine masses but reoccurrence of pyometra due to an impatent cervical canal. The pyometra condition was resolved by insertion of a cervical stent for prevention of intrauterine fluid accumulation. In conclusion, uterine masses, which may severely impact fertility, are best diagnosed by hysteroscopy. Intrauterine purulent concrements should be considered as an atypical form of equine pyometra.     

Comparison of endometrium before and after repair of third-degree rectovestibular lacerations in mares.

The endometrial response of mares to repair of third-degree rectovestibular lacerations was evaluated. Endometrial biopsy specimens from 8 mares with third-degree rectovestibular laceration were obtained immediately before surgery and from 9 to 15 days after repair. Presurgical endometrial biopsy specimens were classified as category I for 2 mares; category II, attributable to slight endometritis, for 5 mares; and category III, attributable to moderate-to-severe endometritis, for 1 mare. Within 15 days after rectovestibular repair, all endometrial biopsy specimens were classified as category I. Results indicated that mares with third-degree rectovestibular injuries are candidates for breeding by artificial insemination by 2 weeks after repair of the injury.     

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.     

Development and Comparison of Sampling Techniques for a Broad Range,Semiquantitative Polymerase Chain Reaction Assay for Detection of Bacterial DNA in the Equine Uterus

Microbial culture from a double-guarded culture swab is commonly used to diagnose infectious endometritis. The objective of this study was to develop a quantitative polymerase chain reaction (qPCR) assay to detect a broad range of bacteria from equine uterine samples. Twenty-seven mares with a clinical history of endometritis had a double-guarded culture swab collected for analysis by qPCR and microbial cultures. An additional 12 mares had a uterine biopsy sample collected for qPCR analysis, microbial culture, and histopathology. Subsequently, a double-guarded culture swab for microbial culture and a cytology brush sample were also collected. The qPCR assay detected bacterial DNA in nine of 27 mares from a double-guarded swab and six of 12 mares from an endometrial biopsy. Positive microbial growth was detected in nine of 27 mares and four of 12 mares from a double-guarded culture swab. Bacterial DNA was detected in two of 27 mares and two of 12 mares without subsequent microbial growth. The simple presence of an organism's DNA allows for detection by nonculture-based systems, both live and dead organisms can be identified. In conclusion, the qPCR assay was determined to be a sensitive diagnostic technique for identifying pathogens associated with infectious endometritis. The primary application of the qPCR assay is detection of potential pathogenic bacteria in the uterus of a mare suspected of having infectious endometritis when a traditional microbial culture is negative. Further work is warranted to determine if mares positive for bacterial DNA and negative for microbial culture are affected clinically.     

Comparison of uterine protein content and distribution of bacteria in the reproductive tract of mares after intrauterine inoculation of Haemophilus equigenitalis or Pseudomonas aeruginosa

Two groups of 3 mares were inoculated with Haemophilus equigenitalis or Pseudomonas aeruginosa on the 1st day of estrus. Uterine flushing samples were recovered on day 3 of estrus and day 8 after ovulation for each cycle. Mares were killed 22, 25, and 30 days after inoculation with P aeruginosa and 45, 46, and 49 days after inoculation with H equigenitalis. Pseudomonas aeruginosa was recovered from the uterus of 2 mares 48 hours after inoculation. Although the initial flushing sample of 1 of these 2 mares had an increased total protein concentration, there appeared to be little difference between protein concentrations of other uterine flushing samples. Haemophilus equigenitalis was recovered from the uterus of each of the 3 mares at postmortem. One mare had a slight, purulent discharge from the vulva. Total protein values were not increased in flushing samples from this mare after inoculation with H equigenitalis. Total protein values decreased in the last flushing sample of each of the 2 remaining mares. Swabbing the uterus was more effective than was homogenizing the uterine mucosa in isolating H equigenitalis.     

Fetal maceration and retention of fetal bones in a mare

A 19-year-old Quarter Horse mare was evaluated because of bloody vaginal discharge that was apparent immediately following breeding. On transrectal ultrasonography, it was evident that the uterus was filled with fluid containing echogenic particles; linear hyperechoic structures were also visible. Endoscopy was performed, which revealed a number of bones adhered to the cranial wall and floor of the right uterine horn as well as purulent fluid in both uterine horns. Bacterial endometritis and fetal maceration were diagnosed. The mare was treated with antibiotics, and the fetal bones were manually removed from the uterus. Fetal maceration with intrauterine retention of bones is rare in mares. Use of hysteroscopy supplements ultrasonography in the diagnosis of uncommon conditions of the uterus. Macerated bones may be adhered to the endometrium, thereby requiring manual removal.     

Morphologic evaluation of acute endometritis in mares with differing resistance to uterine infections

The study was designed to determine differences between normal mares and mares with endometrial pathology in the inflammatory response after bacterial challenge. Six normal mares (biopsy category I) and 4 mares with pathological endometrial changes (biopsy category II) were given an intrauterine infusion of β-hemolytic streptococci on the second day of estrus. All mares had a similar kind of inflammatory response after the bacterial inoculation as assessed by rectal and vaginal examinations. There were no significant differences in the amount of discharge, uterine tone, uterine size and cervical relaxation between the groups. Leukocytic response, as determined by endometrial smears and biopsies, was of the same magnitude in both groups. Two mares from the pathological group were not able to eliminate the infection, but had vaginal discharge and bacteriologically positive uterine swabs until the end of the experiment. It is concluded that the inability of some mares to clear uterine infections cannot be explained by a deficient inflammatory response.     

Feasibility and Safety of Endometrial Injection of Autologous Bone Marrow Mesenchymal Stem Cells in Mares

​Chronic degenerative endometritis (CDE) is an important cause of fertility problems in older mares. It is estimated that 30% of mares breeding are over 18 years old and the high value of their progeny encourages the use of these animals in assisted reproduction procedures. Currently, cell-based therapies are broadly used in human and veterinary regenerative medicine and have been showed a good effect on the treatment of liver fibrosis. Thus, the present study aimed to evaluate the feasibility and safety of endometrial injection of autologous bone marrow MSCs in mares. Mares were examined by transrectal ultrasound and the endometrial stem cells injections performed during diestrous, using injection needles coupled to a teflon catheter through the biopsy channel of a flexible endoscope. After treatment clinical evaluations (heart reat, respiratory rate, staining of the mucosa, capillary refill time, body temperature and lameness score) were performed daily during seven days. The intrauterine fluid and endometrial edema was evaluated before, 24 and 48 hours after the procedure by ultrasonographic exams and the inflammatory infiltrate (polymorphonuclear cells and mononuclear cells) and degree of fibrosis by histological evaluations before (D0) and 15 (D15), 30 (D30) and 60 (D60) days after endometrial injections. The results were evaluated by Kolmogorov-Smirnov and Kruskal-Wallis test followed by Dunn test and for fertility rates was used Chi-square, considering a 5% significance level. Neither clinical alteration was observed in mares after treatment, as well as, intrauterine fluid and endometrial edema were not detected in any mare before and after cell therapy. After cell therapy, significantly more PMNs were found in D15 biopsies, however, these cells were not observed in D30 and D60 biopsies. No worsening on the histological architecture after treatment was observed in any mare. In conclusion, the results of this study showed that endometrial hysteroscopic injections of stem cells in mares is a safe procedure.     

A review of fungal endometritis in the mare

Fungal endometritis is a relatively rare cause of infertility in the mare, accounting for only 1–5% of all cases of endometritis. However, it remains a challenge to the clinician due to difficulties in diagnosing and effectively treating affected mares, resulting in a high rate of recurrence and a guarded prognosis for ongoing fertility. Often occurring as an opportunistic infection, the most common causes of fungal endometritis in the mare are yeasts (Candida spp.) and moulds with septated hyphae (Aspergillus spp.). Early detection and identification of the causative agent are vital when choosing appropriate treatment as sensitivities to commonly used antifungals vary significantly. Assessing in vitro sensitivity for each isolate is accordingly recommended. Treatment of mares with fungal endometritis is multifaceted and includes; correction of any anatomical defects, uterine lavage and systemic and/or intrauterine infusion of antifungals. Concurrent bacterial infection is common so anticipation and subsequent treatment of bacterial endometritis is also often required.     

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding‐induced endometritis and the effect of a specific iNOS inhibitor in vitro

Emerging research suggests that the nitric oxide system may play a role in persistent breeding‐induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose‐dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post‐challenge and cultured at 37°C for 24 hr in L‐arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non‐parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares.     

Inflammatory mechanisms of endometritis

Transient post breeding endometritis is a normal physiological reaction in the mare, as it is believed that an inflammatory response is necessary for the effective removal of contaminating bacteria and excess spermatozoa introduced into the uterus. While most mares can clear endometritis within a reasonable amount of time, persistent endometritis caused by either bacteria or spermatozoa can threaten the success of a pregnancy. A subpopulation of mares is susceptible to persistent endometritis, and these mares are a concern in equine reproductive medicine. Research has identified several factors that contribute to susceptibility; however, the exact mechanisms of the progression of the disease are still being elucidated. Current research focuses on endometrial gene expression during endometritis in an attempt to understand the timing of specific inflammatory processes involved with the development of susceptibility to persistent endometritis. With an increased understanding of the mechanisms involved with the disease, current treatments can be improved upon, and new treatments can be developed to target affected pathways. The Summary is available in Chinese – see Supporting information.     

The technique and clinical application of endometrial cytology in mares

Experimental studies of endometrial smears from 26 normal mares showed that after a period of sexual rest exceeding 7 days smears were generally free from inflammatory cells. Smears from 9 mares showing signs of persistent endometritis contained inflammatory cells in 91 per cent while bacterial culture was positive in only 45 per cent. Smears were also taken from 242 Thoroughbred mares as a routine procedure in 1977. It was concluded that endometrial cytology provides a better guide to uterine inflammation than bacteriology. The routine clinical use of endometrial cytology is recommended to aid the interpretation of both positive and negative bacterial cultures from the cervix.     

Degenerative Endometrial Changes do not Change the Functional Capacity of Immigrating Uterine Neutrophils in Mares

An endometritis model was used to investigate the influence of degenerative endometrial changes (endometrosis) on functional parameters of uterine neutrophils in the horse. Six hours after intrauterine application of recombinant human interleukin‐8 (rhIL‐8), the uteri of 15 mares were flushed with phosphate‐buffered saline. Quantitative and qualitative flow cytometric assays were then made to determine the absolute numbers, viability, phenotype, generation of reactive oxygen species (ROS), and phagocytic activity of immigrated polymorphonuclear neutrophilic granulocytes (PMN). Recombinant hIL‐8 attracted similarly high numbers of similarly viable PMN into the uteri of mares with or without degenerative endometrial changes. Compared with blood PMN, immigrated uterine neutrophils displayed significantly upregulated expression of CD11a/CD18 (LFA‐1) on uterine PMN whereas major histocompatibility complex class I molecules were expressed at lower densities. The ability to phagocytose opsonized streptococci did not differ between uterine and blood PMN. However, uterine PMN displayed a higher capacity to generate ROS. On average, uterine PMN of mares with degenerative endometrial changes showed phenotypical and functional characteristics similar to those of mares with a histologically healthy endometrium. Therefore, degenerative endometrial changes per se did not reduce the functional capacity of equine uterine neutrophils in mares.     

The Effects of Treatment with N-Acetyl Cysteine on Clinical Signs in Persistent Breeding-Induced Endometritis Susceptible Mares

Persistent breeding-induced endometritis (PBIE) is a major cause of infertility in mares. Endometrial inflammation that persists until embryonic descent ultimately results in early embryonic death. A poor endometrial biopsy grade (IIb or III) has been identified as a risk factor for PBIE. Intrauterine fluid accumulation (>2 cm in depth), pathologic endometrial edema, and elevated intrauterine neutrophil levels are all clinical features of PBIE. Commonly applied treatment options include uterine lavage and oxytocin therapy. N-acetyl cysteine (NAC), a mucolytic used to treat bacterial endometritis in mares, has anti-inflammatory properties and was investigated as a potential treatment for PBIE. A randomized, blinded, cross-over design clinical trial used NAC before breeding in PBIE-susceptible mares (n = 9). Intrauterine infusion of 3.3% NAC was performed 12 hours before insemination, and endometrial cytology and endometrial biopsy samples were obtained at 12 and 60 hours after insemination. Endometrial biopsies were evaluated for the degree of inflammation present. Clinical signs of endometrial edema and intrauterine fluid volumes were assessed by transrectal ultrasound at 12 and then every 24 hours after breeding. Data were analyzed using repeated measures analysis of variance and a Mann Whitney Wilcoxon Test. Treatment with NAC did not improve clinical signs in PBIE-affected mares. However, endometrial biopsies from mares treated with NAC displayed more diffuse and severe neutrophil infiltration than control cycles. Further research using a larger population of mares is required to evaluate the effects of NAC treatment on the endometrium of PBIE-susceptible mares.     

Persistent breeding-induced endometritis after hysteroscopic insemination in the mare

Low-dose insemination has been proposed to reduce persistent breeding-induced endometritis (PBIE) in mares with delayed uterine clearance (DUC). Others proposed that hysteroscopic insemination induces an exaggerated inflammatory response and should be avoided in DUC mares. The objectives here were to evaluate presence and severity of PBIE in normal and DUC mares after hysteroscopic insemination with fresh semen, and to determine if hysteroscopy could be used in DUC mares without inducing excessive inflammation. Reproductively normal (n = 4) and DUC (n = 5) mares received four treatments in random order: uterine body insemination (UB, 1 × 10(9) spermatozoa, 20 ml), hysteroscopic insemination (HYST, 5 × 10(6) spermatozoa, 0.5 ml), sham hysteroscopic insemination (SHAM, semen extender, 0.5 ml) and hysteroscopic infusion of seminal plasma (SP, 0.5 ml). Significantly more DUC (50%) mares than normal (14%) mares accumulated intrauterine fluid 24 h post-treatment. The difference in fluid accumulation between DUC (40%) mares and normal (7%) mares was also significant 48 h post-treatment. Fluid scores were not significantly different between treatments in normal mares. However, treatments HYST and SHAM resulted in significantly higher fluid scores 24 h but not 48 h post-treatment in DUC mares. There was no effect of treatment or mare group on the percentage and total number of neutrophils in uterine fluid 48 h post-treatment. Percentage of neutrophils was correlated with duration of hysteroscopy in normal mares, with procedures lasting ≥ 9 min associated with PBIE. There was no effect of mare group, treatment or duration of hysteroscopy on pregnancy rate. Hysteroscopy induces a transient inflammation that is not more severe than that after conventional artificial insemination, suggesting no contraindication to its use in DUC mares.     

SCINTIGRAPHIC MEASUREMENT OF UTERINE CLEARANCE IN MARES

Uterine clearance of technetium 99m-albumin colloid (^99mTc-μAA) was qualitatively and quantitatively measured in 5 reproductively normal mares and 5 mares susceptible to endometritis (infertile). The percentage of 370 MBq ^99mTc-μAA cleared from the uterine lumen within 2 hr of intrauterine infusion was measured in 10 mares on day 3 of estrus and 48 hr after ovulation. The procedure was repeated 3 times on day 3 of estrus in 6 mares to determine repeatability. Six mares were infused with 1110 MBq ^99mTc-μAA on day 3 of estrus to evaluate the effect of increasing the dose to reduce the imaging time. There was no statistically significant difference in the mean percentage of radiocolloid cleared from the uterus during day 3 of estrus or 48 hr after ovulation or in the percent cleared when the studies were repeated in individual mares. There was no statistically significant difference in uterine clearance between the 370 and 1110 MBq dose studies in each mare from 15 to 120 min. Reproductively normal mares cleared approximately 50% of the radiocolloid from the uterus by 120 min while susceptible mares cleared less than 15%.