A tubo-ovarian cyst, paraovarian cysts and lesions of the oviduct in the mare

A tubo-ovarian cyst caused unilateral obstruction of ovum transport in a young Thoroughbred mare. Its effect was a reduction in reproductive efficiency since ovulations from the affected ovary were unproductive. No occluded oviducts were found among 24 pairs of oviducts examined at post mortem. Occlusion of the oviducts in mares is apparently extremely rare. The most common types of cysts found in association with the ovaries in mares are small paraovarian cysts. They are usually found in the fimbriated portion of the oviduct and do not interfere with fertility.     

Heterogenous and xenogenous fertilization of in vivo matured equine oocytes

Forty-five in vivo matured equine oocytes were recovered from 63 follicular aspiration attempts (71.4%). HCG did not improve recovery rate (65% — 24/37 for treated vs 81% — 21/26 for nontreated mares). Fifteen oocytes were transferred into the oviduct of inseminated recipient mares (heterogenous fertilization) and 15 oocytes plus equine spermatozoa were transferred into rabbit oviducts (xenogenous fertilization). Ten oocytes (3 fertilized) were recovered from recipient mare oviducts following removal and flushing two days after transfer. Eight oocytes (nonfertilized) were recovered from rabbit oviducts. Oviductal transfer into separate recipient mares of three embryos produced from heterogenous fertilization resulted in two pregnancies. One mare produced a normal live foal and the other mare aborted at 20 days of gestation. Results from these studies suggest that: 1) a reliable method for collection of in vivo matured oocytes has been established, and 2) heterogenous fertilization is a technique that with refinement should be immediately applicable to obtain foals from valuable infertile mares that fail to get pregnant or produce embryos by standard methods.     

Histological Characteristics of the Equine Oviductal Mucosa at Different Reproductive Stages

The objective of this work was to study cellular changes in the epithelium of the mare’s oviduct. Oviductal samples were taken from mares at different reproductive stages for optical microscopy and Hoechst 33258 staining. Glandular-like structures were observed in 100% of the oviducts. These structures were of the tubular type and were formed by ciliated and nonciliated epithelial cells arranged in a way similar to the epithelial surface. The amount of structures decreased progressively from the ampulla to the isthmus, but did not change through the different reproductive stages. Histological changes in the epithelium of the oviduct were observed associated with the reproductive stages. In the ampulla, the amount of ciliated cells decreased in the anovulatory phase compared with other reproductive stages. Cords of connective tissue lined by epithelium (trabeculae) and dividing completely the lumen of the oviduct were found in 50% of the oviducts. Epithelial cells projected toward the lumen as large vesicles of cytoplasm, sometimes containing a nucleus. The amount of cells presenting nuclear protrusion varied throughout the oviduct, with highest incidence in the ampulla, decreasing progressively toward the isthmus (P < .05). In addition, nuclear protrusions were higher in number during the anovulatory and luteal phases than in the other reproductive stages (P < .05). These nuclear protruding cells appeared to be extruding from the epithelium and showed no signs of apoptosis based on the histological and fluorescent stains used. The existence of these gland-like structures in the oviductal mucosa should be considered when studying the oviductal physiology in mares.     

Isolation and characterization of Chlamydophila psittaci isolated from laying hens with cystic oviducts

The objective of this study was to isolate and identify a hypothetical Chlamydiaceae pathogen from laying hens with an oviduct cyst, and to characterize its potential causal relation with decreased egg production. Our clinical survey showed that cystic oviducts were prevalent at rates of 10% and 15.1% in breeder and commercial hen flocks, respectively. Chlamydial antigens were detected in 20 of 50 pharyngeal swabs (40%) and in 17 of 20 oviduct tissues (85%) using enzyme-linked immunosorbent assay (ELISA) antigen detection kits. The isolated pathogen was identified as Chlamydophila psittaci via complement fixation test, PCE-ELISA, and immunofluorescence assay. Avian influenza virus, Newcastle disease virus, and infectious bronchitis virus were excluded after oviduct tissues were inoculated onto the chorioallantoic membrane of embryonating eggs. The nucleotide sequence of the omp1 gene (accession no. EF202608) from the isolate was similar to that of C. psittaci avian type C (accession no. L25436). Typical cystic oviducts were observed in specific-pathogen-free hens inoculated intraperitoneally with the isolate. The high presence of chlamydial antigen is consistent with the cystic oviducts and poor egg production. We conclude that the isolated C psittaci is most likely associated with cystic oviducts in laying hens.     

Light and electron microscopic studies on mast cells of the bovine oviduct

This study was conducted to determine the staining properties, the light and electron microscopic appearance and numerical distribution of mast cells from different regions of the bovine oviduct during the estrous phase and luteal phase. Ten oviducts from bovines in the estrous phase, and 10 from bovines in the luteal phase were used as study samples. In three regions of the oviduct (infundibulum, ampulla, isthmus), mast cells that showed metachromasia with toluidin blue and Ab(+), SO(-) with the combined Ab/SO stains were encountered. Electron microscopic studies of the mast cells showed them to contain two types of granules; a dense homogeneous type and a tiny particulate type. In all three regions of the oviduct, the luteal phase was observed to have a higher number of mast cells per mm2 area than the estrual phase with the isthmus having the most dense population.     

Laparoscopic application of PGE2 to re-establish oviducal patency and fertility in infertile mares: a preliminary study

REASONS FOR PERFORMING STUDY: Mares are occasionally encountered that consistently fail to conceive when inseminated, naturally or artificially, with fertile stallion semen in the absence of any identifiable pathology of either the structure or function of their reproductive tract. HYPOTHESIS: Temporary blockage of the oviducts by accumulations of naturally occurring oviducal masses may be preventing oviducal transport of the embryo to the uterus. METHODS: Mares, with known reproductive histories, that had exhibited inexplicable failure of conception were treated by laparoscopically guided administration of PGE2-laced triacetin gel directly onto the surface of their oviducts. RESULTS: Fifteen mares age 10-21 years that had exhibited inexplicable failure of conception during 1-4 years were treated, of which 14 (93%) conceived within the same or subsequent breeding season. CONCLUSIONS: The high success rate of this treatment supports the tentative diagnosis of oviducal obstruction in these mares and indicates that blockage of the mare's oviducts may occur in the form of a moveable accumulation of debris rather than from permanent fibrous adhesions resulting from salpingitis. POTENTIAL RELEVANCE: This laparoscopic application of PGE2 to the oviducts constitutes a sound and practical method of restoring fertility in mares suffering oviducal obstruction and further studies involving the procedure are warranted.     

Videoendoscopic evaluation of the mare's uterus: II. Findings in subfertile mares.

Videoendoscopy of the reproductive tract was performed in 87 Thoroughbred mares with histories of reduced fertility. During hysteroscopy samples for cytological, microbiological and histological examinations were obtained under visual control. Common findings in these broodmares included: (a) endometrial degeneration, as assessed by an uneven distribution or atrophy of endometrial folds and/or a scarred appearance of the endometrium (49 mares, 56%); (b) endometrial cysts of various sizes and locations within the uterus with the most common location being at the base of the uterine horns (48 mares, 55%); (c) fluid accumulation in the uterine lumen (28 mares, 32%). A few mares had transluminal adhesions (7 mares, 8%) and in 2 mares the adhesions appeared to obstruct one uterine horn completely. A solitary discrete lump was detected in the wall of the uterine body in one mare and the suspicion of it being a leiomyoma was confirmed histologically with the aid of a visually directed biopsy sample. Free intraluminal structures were present in the uterine lumen in 3 mares, including one inspissated blood clot and two suspected remnants of resorbing pregnancies. Flexible biopsy forceps and scissors passed through the working channel of the endoscope were used to sever small thin adhesions, but this method proved inadequate for multiple adhesions or cysts. Solitary endometrial cysts were removed by means of conventional rigid biopsy forceps passed alongside the endoscope, although bleeding from the operation site usually limited this type of intervention.     

Meiosis Resumption of Canine Oocytes Cultured in the Isolated Oviduct

The aim of this study was to investigate the effects of culture in isolated oviducts relative to meiotic maturation, the time required to resume meiosis and the viability of the canine oocytes. For this purpose, cumulus–oocyte complexes and isthmus–ampullar tracts of the oviducts were collected from bitches undergoing ovariohysterectomies and destined to two experiments of culture. In experiment 1, the oocytes were cultured for 24 or 30 h: (1) in 100 μl drops under oil; (2) on the mucosal epithelium of the open oviducts; (3) in the ligated oviducts. In experiment 2, oocytes were cultured in the ligated oviduct for 24, 30 and 48 h. A group of control oocytes was not cultured (0 h). The results showed that within 30 h of culture, a higher proportion of oocytes (p < 0.001) resumed meiosis in the ligated oviduct (63.8%) than in drop (20.4%) or in the open oviduct (27.1%). Moreover, 24 and 30 h of culture assured higher proportions of meiosis resumption than 48 h (69.2 and 59.1% vs 35.8%, p < 0.005). Oocyte resumption of meiosis was mainly determined by oocytes at meiotic stages preceding metaphase I, while stages between metaphase I and II in the ligated oviduct ranged between 12.5 and 31.9%. The extension of the culture time up to 48 h in the oviduct increased oocyte degeneration significantly (59.3%, p < 0.0001) compared with 24 and 30 h (18.7 and 27.3%, respectively) and the oviductal epithelium showed nuclear picnosis and degeneration following culture. The present study suggests that the close physical interaction between the canine oocytes and the oviductal tract positively affects oocyte maturation, and meiosis is resumed within 30 h of culture. Moreover, the oocyte survival is better preserved within 30 h in the ligated oviduct compared with the conventional culture in drop or to the culture in the open oviduct, but the ligated oviduct does not assure viability of the oocytes up to 48 h of culture.     

Accumulation of Fluid in the Infundibulum During the Estrous Cycle in Mares

The funnel-shaped cranial portion (infundibulum) of the oviduct is contiguous with the ovulation fossa in mares. An accumulation of fluid in the infundibular area was discovered by transrectal ultrasonic imaging and was studied daily in both oviducts of 12 mares from day –10 to day 10 (day 0 = ovulation), and from day –6 to day 6 during 35 estrous cycles of young, intermediate, and old mares (n = 8 mares/group). The infundibulum was identified by processes (fimbriae) and folds in the pocket of fluid. The amount of fluid accumulation was scored from 0 to 3 (nil to maximum). Frequency of detection of fluid in the infundibular area increased between day –10 (46% of oviducts) and day –3 (88%), and decreased between day –3 and day 7 (8%; P < .002). The day-to-day profile for changes in the score for amount of fluid was significant (P < .0001) and similar to the profile for frequency of detection of the infundibulum. The profiles for the two infundibular end points and scores for endometrial echotexture (an indicator of edema) were similar to the reported profile for systemic estradiol concentrations. The frequency of infundibulum detection was greater (P < .0009) for the side ipsilateral to the preovulatory follicle and ovulation (51%) than for the opposite side (36%). No difference among ages was found for either oviductal end point. Results indicated that changes in the amount of fluid accumulation in the infundibular area and endometrial edema were estrous cycle dependent and similar to previously reported changes in systemic concentrations of estradiol.     

Distribution of spermatozoa in the genital tract of artificially inseminated heifers

Eight heifers were artificially inseminated in the uterine body with 160×10⁶ spermatozoa frozen in French mini-straws. The heifers were slaughtered 2 (n = 4) or 12 (n = 4) h after insemination and spermatozoa were recovered by flushing defined segments of the reproductive tract. The efficiency of the method was checked in different ways. There was a slight underestimation of the number of recovered spermatozoa. This underestimation was randomly distributed among heifers and genital tract segments.The total number of spermatozoa recovered was higher at 2 than at 12 h (14.6 vs 0.6 % of the total number inseminated). Most spermatozoa were found in the vagina both at 2 and 12 h after insemination and in greater number at 2 h. In uterus there was a slight decline in the number of spermatozoa recovered at 2 versus 12 h after insemination. The number of spermatozoa recovered from the oviducts were similar at 2 (89.6 × 10³) and 12 h (71.5 × 10³) after insemination. At 2 h spermatozoa were found in all parts of the oviduct with the majority located in the utero tubal junction, whereas at 12 h the most were recovered from isthmus.More spermatozoa were recovered from the left than from the right side of the tract in 6 of the 8 heifers. Only in 1 heifer were the majority of spermatozoa found in the oviduct ipsilateral to the follicle bearing ovary.     

Sperm localization in the oviducts of artificially inseminated dairy cattle

Eight animals, 3 heifers and 5 primiparous cows, were artificially inseminated by intrauterine deposition of frozen-thawed semen. The insemination dose comprised 20×10⁶ or 200 × 10⁶ spermatozoa, frozen in French mini straws. Four animals were inseminated at fixed time interval (72 or 84 h) after cloprostenol injection. The remaining 4 animals were inseminated in spontaneous oestrus. Slaughter took place 2 or 12 h after insemination. After fixation the oviducts were cut into segments, which were serial-sectioned and stained. Six sections per segment were examined under the microscope for sperm recovery.The number of spermatozoa recovered from the oviducts varied considerably among animals. Recovery was poor (less than 50 spermatozoa) in 4 animals. Recovery was low when insemination took place in induced oestrus and with the lower sperm number (20×10⁶). In animals in which more than 50 spermatozoa were found the distribution varied both between animals and between oviducts within the same animal. Overall, more spermatozoa were found in the lower (UTJ, isthmus and AIJ) than in the upper (ampulla) parts of the oviducts. In 3 out of 4 animals more spermatozoa were recovered from the left than from the right oviduct. Only in 1 animal were the majority of spermatozoa found in the oviduct ipsilateral to the follicle-bearing ovary.     

Fertility of mares after unilateral laparoscopic tubal ligation

OBJECTIVE: To develop a technique for laparoscopic tubal (oviductal) ligation and to evaluate pregnancy rates for mares that ovulated ipsilateral or contralateral to the ligated oviduct. STUDY DESIGN: Randomized prospective clinical trial comparing pregnancy rates after unilateral laparoscopic tubal ligation. ANIMALS: Twelve mares of light horse breeds. METHODS: One oviduct in each of 6 mares was surgically ligated with a laparoscopic technique; 6 other mares served as nonligated controls. Mares with unilateral tubal ligations (UTL) were inseminated with 500 million progressively motile sperm during 1 cycle when the dominant follicle was ipsilateral to the ligation site and 1 cycle when the dominant follicle was contralateral to the ligation site. Control mares were bred during 2 cycles regardless of the side of the dominant follicle. Pregnancy examinations were performed on days 12, 14, and 16 after ovulation by transrectal ultrasonography. RESULTS: None of the mares became pregnant when ovulations occurred from the ovary adjacent to the ligated oviduct. All 6 mares became pregnant on the first cycle when an ovulation occurred from the opposite ovary. Control mares became pregnant on 10 of 12 cycles (83.3 %). CONCLUSIONS: UTL was completely effective in preventing pregnancy when ovulation occurred ipsilateral to the ligation site. The surgical procedure did not interfere with the establishment of pregnancy when ovulation occurred from the contralateral ovary. CLINICAL RELEVANCE: UTL may be a clinically useful procedure for preparing a recipient mare for gamete intrafallopian transfer. The recipient mare could be allowed to ovulate and UTL would prevent fertilization of her oocyte but would not interfere with normal corpus luteum formation. The donor oocyte could be placed into the oviduct contralateral to the UTL site.     

Electrocoagulative removal of endometrial cysts in the mare

Six mares had endometrial cysts removed by endoscopic electrocoagulative surgery. These mares exhibited poor reproductive histories and endometrial cysts were the only detectable abnormalities. In each case, at least two cysts greater than 2.5cm were present. One mare subsequently produced a live foal; two are presently in foal; one conceived but absorbed; one formed more cysts after surgery; and one has not yet been bred. Histologically, the walls of the cysts contained entire lamina propria, relatively normal endometrial glands, and small cystic spaces.     

Characterisation of lymphocyte subsets in the equine oviduct

REASONS FOR PERFORMING STUDY: The equine oviduct is the site of fertilisation and location of embryonic development during the first 5 or 6 days. It therefore has an important influence on mare fertility. Although histopathological changes have been described previously, there is limited information regarding lymphocyte subtypes present in the mucosa of the normal equine oviduct. OBJECTIVES: To characterise the distribution of CD3+, CD4+, CD8+ and B lymphocytes in the equine oviduct from inseminated mares during oestrus and dioestrus, and from noninseminated mares during the immediate post ovulatory period. METHODS: Oviductal tissues were collected from noninseminated mares at oestrus (> 30 mm follicle, n = 4), at Day 1 post ovulation (n = 3) and at dioestrus (Day 7 post ovulation; n = 4). Oviducts were also collected from inseminated mares at Days 1, 2, and 3 post ovulation (n = 4 for each period). Cross-sections of tissues from the ampullar-isthmic junction from each oviduct were snap frozen and cryostat sections stained by the immunoperoxidase technique with monoclonal antibodies directed against equine lymphocyte surface markers for B cells as well as CD3+, CD4+ and CD8+ cells. RESULTS: In all oviductal sections examined, B cells were rare whereas T cells were relatively abundant. The predominant cell type found was the CD8+ phenotype, with a lesser number of CD4+ cells. Among mares, individual variation was large; therefore, although breeding status and stage of oestrous cycle appeared to alter lymphocyte populations, these differences were not significant. CONCLUSIONS AND POTENTIAL RELEVANCE: A population of CD3+, CD4+ and CD8+ cells exists within the mucosal region of the equine oviduct. The density of these cells is similar to that described in the human oviduct. Their function is not currently known, but they may be involved with modulation of the maternal response to the presence of spermatozoa or the early conceptus within the equine oviduct. As our capacity to differentiate these cell types improves, along with the ability to identify the specific cytokines they produce, their functional significance will become more apparent.     

Cyclic Related and Pathological Changes in the Lectin-binding Sites on the Swine Oviduct

This study was carried out to compare the lectin-binding pattern in the normal and pathological oviduct of sows during the ovarian cycle. Lectin-binding patterns showed differences between segments, phases of ovarian cycle and presence of morphologic lesions. In the infundibulum, it was observed that the cysts, in the follicular phase, reduced Ricinus communis-I (RCA-I) and Ulex europaeus-I (UEA-I) binding. Furthermore, in the pathological oviducts of the luteal-phase group, there is a reduction of Concanavalia ensiformis (Con-A) reactivity in this segment of the tube having wall cysts, adenomyosis and diverticulus. The Arachis hypogaea (PNA) binding in the infundibulum, during the luteal phase, decreased in the tube having adenomyosis. In animals with wall cysts, the Con-A, Triticum vulgaris (WGA) and RCA-I reactivity was minor in the glycocalyx of the isthmus epithelium during follicular phase. Con-A and Dolichos biflorus (DBA) binding pattern was minor in the luteal-phase isthmus of the tube having wall cysts, adenomyosis and diverticulus. In the ampulla, the wall cysts impaired the Con-A reaction only in the basal region of the epithelium, in the follicular phase. Binding with Con-A was decreased in the ampulla of animals in the luteal phase in the tube lesions with cysts and diverticulus. In addition, the diverticulus observed in the ampulla, during the luteal phase, reduced the PNA tubaric binding. The results of this study showed that the morphologic alterations modify the sugar pattern in the oviduct of sows. These modifications in glycoconjugates may be one of the reasons for the failure of fertility in sows.     

Effect of eFSH on Ovarian Cyclicity and Embryo Production of Mares in Spring Transitional Phase

The use of equine FSH (eFSH) for inducing follicular development and ovulation in transitional mares was evaluated. Twenty-seven mares, from 3 to 15 years of age, were examined during the months of August and September 2004, in Brazil. Ultrasound evaluations were performed during 2 weeks before the start of the experiment to confirm transitional characteristics (no follicles larger than 25 mm and no corpus luteum [CL] present). After this period, as the mares obtained a follicle of at least 25 mm, they were assigned to one of two groups: (1) control group, untreated; (2) treated with 12.5 mg eFSH, 2 times per day, until at least half of all follicles larger than 30 mm had reached 35 mm. Follicular activity of all mares was monitored. When most of the follicles from treated mares and a single follicle from control mares acquired a preovulatory size (≥35 mm), 2,500 IU human chorionic gonadotropin (hCG) was administered IV to induce ovulation. After hCG administration, the mares were inseminated with fresh semen every other day until ovulation. Ultrasound examinations continued until detection of the last ovulation, and embryo recovery was performed 7 to 8 days after ovulation. The mares of the treated group reached the first preovulatory follicle (4.1 ± 1.0 vs 14.9 ± 10.8 days) and ovulated before untreated mares (6.6 ± 1.2 vs 18.0 ± 11.1 days; P < .05). All mares were treated with prostaglandin F_2α (PGF_2α), on the day of embryo flushing. Three superovulated mares did not cycle immediately after PGF_2α treatment, and consequently had a longer interovulatory interval (22.4 vs 10.9 days, P < 0.05). The mean period of treatment was 4.79 ± 1.07 days and 85.71% of mares had multiple ovulations. The number of ovulations (5.6 vs 1.0) and embryos (2.0 vs 0.7) per mare were higher (P < 0.05) for treated mares than control mares. In conclusion, treatment with eFSH was effective in hastening the onset of the breeding season, inducing multiple ovulations, and increasing embryo production in transitional mares. This is the first report showing the use of FSH treatment to recover embryos from the first cycle of the year.     

Passive hemagglutination test for detection of antibodies against Taylorella (Haemophilus) equigenitalis in sera of mares

The passive hemagglutination (PHA) test was improved to enable the detection of antibodies to Taylorella (Haemophilus) equigenitalis in the sera of mares. Horse red blood cells (RBC) fixed with glutaraldehyde were compared with similarly treated RBC of a cow, pig and sheep for the PHA test. The horse RBC were superior to those of the other animals tested in detecting mares affected with contagious equine metritis (CEM). A PHA test using these cells as indicator and an antigen prepared from T. equigenitalis by sonication following treatment with hyaluronidase was the most satisfactory in terms of sensitivity and specificity. None of the 156 serum samples from clinically healthy mares without a history of contact with T. equigenitalis-infected stallions or mares showed PHA titers greater than 1:32 and only a few samples (7.1%) showed PHA titers of 1:32. Four of the 50 serum samples from mares affected with CEM showed PHA titers of 1:32, while most of the samples (92.0%) showed PHA titers greater than 1:32. The glutaraldehyde-fixed horse RBC sensitized with the antigen had the advantage of being reproducible for at least 7 months when preserved at 4 degrees C.     

Use of oocyte transfer in a commercial breeding program for mares with reproductive abnormalities

In some mares with lesions of the reproductive tract, embryo collection and survival rates are low, or collection of embryos is not feasible. For these mares, oocyte transfer has been proposed as a method to induce pregnancies. In this report, a method for oocyte transfer in mares and results of oocyte transfer performed over 2 breeding seasons, using mares with long histories of subfertility and various reproductive lesions, are described. Human chorionic gonadotropin or an implant containing a gonadotropin-releasing hormone analog was used to initiate follicular and oocyte maturation. Oocytes were collected by means of transvaginal ultrasound-guided follicular aspiration. Following follicular aspiration, cumulus oocyte complexes were evaluated for cumulus expansion and signs of atresia; immature oocytes were cultured in vitro to allow maturation. The recipient's ovary and uterine tube (oviduct) were exposed through a flank laparotomy with the horse standing, and the oocyte was slowly deposited within the oviduct. Oocyte transfer was attempted in 38 mares between 9 and 30 years old during 2 successive breeding seasons. All mares had a history of reproductive failure while in breeding and embryo transfer programs. Twenty pregnancies were induced. Fourteen of the pregnant mares delivered live foals. Results suggest that oocyte transfer can be a successful method for inducing pregnancy in subfertile mares in a commercial setting.     

Interaction of Tritrichomonas foetus and the bovine oviduct in an organ culture model

Tritrichomonas foetus is an extracellular parasite of the reproductive tract in cattle. The mechanism by which T. foetus causes abortion in cattle is largely unknown. There are no studies of infection in the cow oviducts, almost all published papers are related to vagina infection and few articles focusing on the uterus. The aim of the present study was to establish a working model of bovine oviduct epithelial cells and submit these cells to Tritrichomonas foetus interaction. Twenty bovine oviducts were obtained from cows at a commercial abattoir and T. foetus was injected through the isthmus into the oviduct lumen. The whole oviduct was analyzed by scanning and transmission electron microscopy. The results reported here demonstrate that: (1) fresh whole oviducts can be used as a good model to study parasite-host cell interaction; (2) cow oviduct epithelium has been shown to consist of two cell types: ciliated and nonciliated secretory cells, and T. foetus displayed great specificity for the nonciliated cells localized in the deeper oviduct folds; (3) T. foetus adheres as single separate cells, and maintains the flagella externalized; (4) differently from T. vaginalis, T. foetus does not change its shape during the adhesion process; and (5) oviduct cells exhibited morphological characteristics of apoptosis after trichomonadal interaction.