Recognizing uterine torsion as a differential diagnosis in pregnant cats with severe anemia to provide appropriate and timely care in the absence of a definitive presurgical diagnosis.

A pregnant female domestic longhair cat ~8 mo of age was referred to the Western College of Veterinary Medicine (Saskatoon, Saskatchewan) for a diagnostic evaluation of severe anemia (PCV: 10.8%) after a 2-day period of lethargy. A CBC, serum biochemistry profile, FeLV/FIV testing, and abdominal radiographs were completed and did not determine a cause for the anemia. Abdominal ultrasonography identified 1 viable and 6 nonviable and fetuses, anechoic fluid in the uterus, and a mild volume of peritoneal effusion. A whole-blood transfusion and C-section with ovariohysterectomy were performed even though a definitive presurgical diagnosis for the anemia had not yet been established. Exploratory surgery revealed a left uterine horn torsion with a necrotic base, severe congestion, and 7 nonviable fetuses. Following surgery, the queen made a full clinical recovery. Key clinical message: Uterine torsion can be easily overlooked as a cause of severe anemia due to the relative infrequency of this condition in cats and the low sensitivity of ultrasonography to provide a definitive presurgical diagnosis. Client communication must emphasize the need for a prompt surgical intervention to establish the diagnosis and to save the cat, despite poor rates of neonatal survival. Once the animal is stabilized after surgery, further diagnostic tests and procedures are indicated if the cause of anemia has not yet been identified.

Reconnaître la torsion utérine comme un diagnostic différentiel chez les chattes gestantes souffrant d'anémie sévère afin de fournir des soins appropriés et opportuns en l'absence d'un diagnostic pré-chirurgical définitif. Une chatte domestique à poils longs, âgée d'environ 8 mois, a été référée au Western College of Veterinary Medicine (Saskatoon, Saskatchewan) pour une évaluation diagnostique d'anémie sévère (hématocrite : 10,8 %) après une période de léthargie de 2 jours. Une formule sanguine complète, un profil biochimique sérique, des tests FeLV/FIV et des radiographies abdominales ont été réalisés et n'ont pas permis de déterminer la cause de l'anémie. L'échographie abdominale a identifié 1 foetus viable et 6 non viables, du liquide anéchoïque dans l'utérus et un léger volume d'épanchement péritonéal. Une transfusion de sang total et une césarienne avec ovariohystérectomie ont été réalisées même si le diagnostic pré-chirurgical définitif de l'anémie n'avait pas encore été établi. La chirurgie exploratoire a révélé une torsion de la corne utérine gauche avec une base nécrotique, une congestion sévère et 7 foetus non viables. Après l'opération, la chatte s'est complètement rétablie cliniquement.Message clinique clé:La torsion utérine peut facilement être négligée comme cause d'anémie sévère en raison de la rareté relative de cette affection chez le chat et de la faible sensibilité de l'échographie pour fournir un diagnostic pré-chirurgical définitif. La communication avec le client doit souligner la nécessité d'une intervention chirurgicale rapide pour établir le diagnostic et sauver le chat, malgré de faibles taux de survie néonatale. Une fois l'animal stabilisé après la chirurgie, d'autres tests et procédures de diagnostic sont indiqués si la cause de l'anémie n'a pas encore été identifiée.(Traduit par Dr Serge Messier).

Expression of aldose reductase in mouse endometrial epithelial cells and its role in sperm capacitation.

The survival, motility and capacitation of sperm in the female reproductive tract are important prerequisites for fertilization. The uterus is the main location for sperm capacitation. One of the most important physiological functions of the endometrial epithelium is to create a suitable uterine environment under the regulation of ovarian hormones, to ensure sperm capacitation. The composition of uterine fluid directly affects sperm capacitation. Fructose is an important component of semen that supports sperm viability and motility. Aldose reductase, a rate-limiting enzyme in the polyol pathway, metabolizes sorbitol and fructose, thereby supplying cells with necessary energy for functional activities. Existing studies have reported the presence aldose reductase in the endometrium, leading us to hypothesize that its expression in endometrial epithelium might promote sperm capacitation by maintaining the uterine environment. Yet, the mechanism of regulation has not been clarified. In this study, we investigated the expression of aldose reductase in mouse endometrial epithelium and its potential role in sperm capacitation. We initially investigated the periodic characteristics of glucose, fructose and sorbitol in uterine fluid. We then studied the temporal and spatial characteristics of aldose reductase in the endometrial epithelium. Next, we examined the effect of aldose reductase on glucose, fructose and sorbitol in uterine fluid. Finally, we explored the effect of aldose reductase on sperm capacitation and fertilization. The results showed that glucose and fructose content in uterine fluid and the expression of aldose reductase fluctuated periodically during physiological periods. Inhibition of aldose reductase in the endometrial epithelium interfered with sperm capacitation and fertilization by reducing the fructose levels in the uterine fluid. To conclude, the aldose reductase-mediated polyol pathway in endometrial epithelial cells is essential to maintain an appropriate fructose environment in the uterine fluid for sperm capacitation and fertilization.

Ultrasonographic diagnosis of adhesions, quantification of uterine wall and managerial perspectives of uterine torsion in buffaloes.

The present study aimed to diagnose uterine adhesion using ultrasonography to decide the treatment approach for better survival of buffaloes suffering from uterine torsion. The peculiar visceral slide against the other abdominal organs induced by breathing was considered an absence of uterine adhesion, while the total absence of visceral slide was considered the presence of uterine adhesions on ultrasonographic examination. The 59 Mehsana buffaloes suffering from uterine torsion were divided into two groups based on the presence (Group I; n = 27) or absence (group II; n = 32) of adhesions. The mean pixel values (MPV) and thickness of uterine wall were also quantified in both the groups. In group I, severe type adhesions (n = 16/27) were directly subjected to caesarean section (CS), while fibrinous adhesions (n = 11/27) were subject to rolling by Sharma's modified Schaffer's method, and if rolling failed were further, subjected to CS and adhesions were confirm during CS. In Group II, 28 buffaloes were subjected to rolling by Sharma's modified Schaffer's method and 4 buffaloes were directly subjected to CS because of longer duration of illness (≥3 days), higher degree (≥180°) and previous field handling. The incidence of adhesions was 45.76 per cent (27/59) in buffaloes with uterine torsion. The degenerative changes such as oedematous (hyperoedema-hypoechoic with anechoic trabeculae) or thin (homogenous hypoechoic), sloughing of serosa and wall separation were observed in the uterine wall. The significantly thinner uterine wall (p = .017) and higher MPV (p = .001) were evident in group I than in group II. The weak negative non-significant correlation between uterine wall thickness and MPV was observed in group I (r = -.250, p = .147) and group II (r = -.235, p = .089). The dam survival rate was 48.15 per cent (13/27) in group I and 81.25% (26/32) in group II. There was a significant (p = .04) association between the treatment approach and dam survival. Ultrasonography has synergistic diagnostic value for uterine adhesions by evaluating uterine surfaces beyond the reach of obstetricians during a manual clinical examination. Thereby, avoiding futile efforts to relieve uterine torsion by rolling might help to improve dam survival by planning a better obstetrical manoeuvre to salvage buffaloes suffering from uterine torsion.

Uterine Inflammatory Response After Prostaglandin E1 (Misoprostol) Infusion Prebreeding or Immediately After Embryo Flushing in Commercial Donor Mares.

Misoprostol, a synthetic PGE1, is becoming a common therapy for mares with suspected uterine tube obstruction. Recently, there have been concerns that uterine administration of misoprostol induces exacerbated uterine inflammation; however, this has not been critically evaluated. This study aimed to assess the inflammatory response and potential systemic reactions after uterine administration of misoprostol, either during prebreeding or immediately after postembryo flushing. Privately owned embryo donor mares (n = 11) were randomly assigned in a crossover design to receive misoprostol (3 mL +200 µg) or sham (3 mL of lactate Ringer's solution) infusions, bilaterally deposited via deep-horn, at least 72 hours prebreeding (experiment 1) or immediately after embryo flushing (experiment 2). Each mare had one cycle for misoprostol and sham in both experiments and a breeding cycle (no sham or misoprostol) between experiments. Uterine edema, fluid accumulation, and the number of uterine PMN were assessed before each infusion and then daily for 72 hours. Uterine lavage was performed the day after each infusion across groups and experiments. Ovulation was hastened with a GnRH agonist and confirmed at 24 hour-intervals. Mares were bred with semen from one of six stallions per owner's choice. Embryo flushing was performed 8 to 9 days postovulation. In either experiment, misoprostol did not affect uterine edema or fluid accumulation (P > .05). However, both the sham and misoprostol infusions increased the number of PMN up to 48 hours postinfusion in both experiments. Embryo recoveries were similar between sham (45%, 5/11) and misoprostol cycles in experiments 1 (45%, 5/11; P > .05) and 2 (sham, 68%, 7/11; misoprostol, 45%, 5/11; P > .05). In conclusion, misoprostol did not induce exacerbated uterine inflammation in mares or systemic adverse reactions when infused prebreeding or immediately after embryo flushing.

[Uterine serosal inclusion cysts in the dog - case report and literature review of canine uterine cysts]. / Serosazysten am Uterus der Hündin ­ Fallbericht und Literaturübersicht zu kaninen Uteruszysten.

In bitches, uterine cysts are usually associated with pathological conditions. Serosal inclusion cysts of the uterus represent an exception to this rule. These thin-walled cysts develop on the uterine surface and occur either as single or multiple cysts. The cysts lack endocrine activity and usually do not cause clinical symptoms. Their occurrence shows no association to the estrus cycle. Serosal inclusion cysts are mainly found in older multiparous bitches and large breeds are more often affected. Diagnosis of serosal inclusion cysts may be achieved by transabdominal sonography. Differentiation between uterine and ovarian serosal inclusion cysts is possibly challenging as serosal inclusion cysts are often located in the area of the tip of the uterine horn. This report presents the case of a breeding bitch with multiple uterine serosal inclusion cysts detected during caesarean section. The concentrations of estradiol 17ß and progesterone in the cyst fluid corresponded to the blood concentrations of these hormones in a bitch during parturition. The concentrations of total calcium, zinc, copper, and iron in the cyst fluid were below the reference values in canine blood whereas the concentrations of magnesium, phosphorus, sodium, and potassium in the cyst fluid were comparable to pertaining blood levels.

The role of somatostatin and its receptors (sstr2, sstr5) in the contractility of gilt inflamed uterus.

We studied the inflammation effect on somatostatin receptors subtypes 2 (sstr2) and 5 (sstr5) expression in myometrium and somatostatin influence alone or with sstr2 and sstr5 antagonists on the contractility of gilt inflamed uterus. On day 3 of the estrous cycle, either E.coli suspension (E.coli group) or saline (SAL group) were injected into uterine horns. In the control pigs (CON group), only laparotomy was performed. Eight days later, in the E.coli group developed severe acute endometritis. In this group, myometrial sstr2 mRNA expression lowered and protein expression increased compared to other groups. Compared to period before somatostatin administration, somatostatin did not change tension in myometrium and endometrium/myometrium of three groups, reduced amplitude and frequency in the CON and SAL groups, and increased amplitude and decreased frequency in the E.coli group. In this group, amplitude was increased by somatostatin compared to other groups. In the CON and SAL groups, sstr2 eliminated inhibitory somatostatin effect on amplitude, while sstr5 antagonist reversed inhibitory somatostatin effect on amplitude. In the E.coli group, sstr2 antagonist reversed stimulatory somatostatin effect on amplitude, while in sstr5 antagonist presence stimulatory somatostatin effect was more deepened compared to somatostatin action alone. After using sstr2 antagonist more deepened inhibitory somatostatin effect on frequency in the CON and E.coli groups was found. Sstr5 antagonist partly eliminated inhibitory somatostatin effect on frequency in the SAL group. Summarizing, the uterine inflammation increases the myometrial sstr2 protein expression; somatostatin raises amplitude of the inflamed uterus acting by sstr2, while drops this parameter by sstr5.

Qualitative and quantitative differences in endometrial inflammatory gene expression precede the development of bovine uterine disease.

The transcriptome of the endometrium early postpartum was profiled to determine if inflammatory gene expression was elevated in cows which subsequently developed uterine disease. Endometrial cytobrush samples were collected at 7 days postpartum (DPP) from 112 Holstein-Friesian dairy cows, from which 27 were retrospectively chosen for RNA-seq on the basis of disease classification [ten healthy and an additional 17 diagnosed with cytological endometritis (CYTO), or purulent vaginal discharge (PVD)] at 21 DPP. 297 genes were significantly differentially expressed between cows that remained healthy versus those that subsequently developed PVD, including IL1A and IL1B (adjusted p < 0.05). In contrast, only 3 genes were significantly differentially expressed in cows which subsequently developed CYTO. Accounting for the early physiological inflammatory status present in cows which do not develop disease enhanced the detection of differentially expressed genes associated with CYTO and further expression profiling in 51 additional cows showed upregulation of multiple immune genes, including IL1A, IL1B and TNFA. Despite the expected heterogeneity associated with natural infection, enhanced activation of the inflammatory response is likely a key contributory feature of both PVD and CYTO development. Prognostic biomarkers of uterine disease would be particularly valuable for seasonal-based dairy systems where any delay to conception undermines sustainability.

Ovarian function and the establishment and maintenance of pregnancy in dairy cows with and without evidence of postpartum uterine disease.

Uterine disease early postpartum reduces fertility during the breeding period. One potential mechanism involves the reduced functional capacity of the uterus to support pregnancy. A second potential mechanism involves damage to ovarian follicles associated with systemic inflammation. We categorized lactating Holstein cows into healthy (n = 63) and diseased (n = 39) uterus groups based on the percentage of polymorphonuclear neutrophils in the uterine lumen during the second and third month postpartum and evaluated the functionality of their ovaries and their capacity to establish and maintain pregnancy. Cows were enrolled in a timed artificial insemination protocol (Presynch Ovsynch) so that the first artificial insemination was approximately 75 d postpartum. Ovarian follicles and corpora lutea were counted and measured using transrectal ultrasound, ovulatory responses were assessed, and luteal phase progesterone concentrations were measured. Pregnancy was detected on d 18, 20, 22, 25, 32, and 45 through chemical (d 18 to 25) or ultrasonographic methods (d 32 and 45). The percentage of cows ovulating during the Presynch period; the number, diameter, and ovulatory capacity of follicles during the Ovsynch period; and plasma progesterone concentrations following ovulation were similar for healthy and diseased cows. The initial period of pregnancy establishment (d 18 to 22) appeared to be unaffected by disease because a similar percentage of healthy and diseased cows were pregnant during this period. Embryonic loss occurred in both healthy and diseased cows after d 22. Based on a relatively small number of pregnancies (n = 30 healthy and n = 17 diseased), the cumulative embryonic loss after d 22 was greater in diseased compared with healthy cows. In short, uterine disease as defined in this study did not affect cyclicity, ovarian follicular growth, or plasma progesterone concentrations. Percentages of healthy and diseased cows that were pregnant were similar from d 18 to 22 after artificial insemination. Greater embryonic loss was observed after d 22 in diseased compared with healthy cows, but this observation was based on a small number of pregnancies and should be studied further in larger trials with greater statistical power.

Lysophosphatidic acid as a regulator of endometrial connective tissue growth factor and prostaglandin secretion during estrous cycle and endometrosis in the mare.

BACKGROUND: Equine endometrosis is a chronic degenerative condition, described as endometrial fibrosis that forms in the stroma, under the basement membrane and around the endometrial glands. The role of lysophosphatidic acid (LPA) in the development of tissue fibrosis varies depending on the organ, and its profibrotic role in mare endometrosis remains unclear. The study aimed to establish the endometrial presence of LPA and its receptors (LPAR1-4), together with its effects on connective tissue growth factor (CTGF) and prostaglandins (PG) secretion from equine endometrium under physiological (estrous cycle), or pathological conditions (endometrosis). Mare endometria in the mid-luteal phase (n = 5 for each category I, IIA, IIB, III of Kenney and Doig) and in the follicular phase (n = 5 for each category I, IIA, III and n = 4 for IIB) were used. In experiment 1, the levels of LPA, LPAR1-4 mRNA level and protein abundance were investigated in endometria at different stages of endometrosis. In experiment 2, the in vitro effect of LPA (10- 9 M) on the secretion of CTGF and PGs from endometrial tissue explants at different stages of endometrosis were determined. RESULTS: Endometrial LPA concentration was higher in the mid-luteal phase compared to the follicular phase in category I endometrium (P < 0.01). There was an alteration in endometrial concentrations of LPA and LPAR1-4 protein abundance in the follicular phase at different stages of endometrosis (P < 0.05). Additionally, LPA increased the secretion of PGE2 from category I endometrium in both phases of the estrous cycle (P < 0.05). The effect of LPA on the secretion of CTGF and PGF2α from endometrial tissue was altered depending on different stages of endometrosis (P < 0.05). CONCLUSION: Our data indicate that endometrosis disturbs proper endometrial function and is associated with altered endometrial LPA concentration, its receptor expression and protein abundance, PGE2/PGF2α ratio, and CTGF secretion in response to LPA. These changes could influence several physiological events occurring in endometrium in mare during estrous cycle and early pregnancy.

Peptidoglycan disrupts early embryo-maternal crosstalk via suppression of ISGs expression induced by interferon-tau in the bovine endometrium.

Uterine infection with bacteria and the release of peptidoglycan (PGN), antigenic cell wall components of both Gram-negative and Gram-positive bacteria, can cause early pregnancy losses in ruminants, but the associated mechanisms remain unsolved. Day 7 blastocyst starts to secrete a minute amount of interferon-tau (IFNT) in the uterine horn which is required for early stage of maternal recognition of pregnancy (MRP) in ruminants, and it induces interferon stimulated genes (ISGs) for driving uterine receptivity in cows. This study investigated if PGN disrupts IFNT response through modulation of endometrial ISGs expressions. Cultured bovine endometrial epithelial cells (BEECs) were treated with embryo culture medium (ECM) or IFNT (1 ng/ml) in the presence or absence of a low level of PGN (10 pg/ml) for 24 h. A real-time PCR analyses revealed that the presence of PGN suppressed IFNT-induced ISGs (OAS1 and ISG15) and STAT1 expressions in BEECs. To visualize the impact of PGN in an ex-vivo model that resembles the in vivo status, endometrial explants were treated by IFNT (1 ng/ml) with or without PGN (10 pg/ml) for 12 h. PGN suppressed IFNT-induced gene expressions of the above factors, but not for IFNA receptor type1 (IFNAR1) or type2 (IFNAR2) in explants. Immunofluorescence analysis illustrated that PGN completely suppressed the IFNT-triggered OAS1 protein expression in the luminal epithelium of explants. Of note, PGN did not stimulate pro-inflammatory cytokines (TNFA and IL1B) or TLR2 mRNA expression in both models. These findings indicate that the presence of low levels of PGN suppresses ISGs expression induced by IFNT secreted from early embryo, at the luminal epithelium of the bovine endometrium. This could severely interfere with early stage of MRP processes in cows, leading to pregnancy failure.

Influence of uterine torsion in water buffaloes (Bubalus bubalis) with insights into the hematological and biochemical prognostic values regarding to manual correction.

Bovine uterine torsion refers to the rotation of the gravid uterus around its longitudinal axis. It is the dominant cause of dystocia in buffaloes. A total of two hundred and seventeen dystocia's Egyptian buffaloes were examined, and another twenty were apparently normal at the full term (healthy pregnant group). From the dystocia's buffaloes, twenty-three percentage (n = 50) was uterine torsion after gynecology inspection at presentation (Ut group) and 5 days after the manual correction (Utc group). The buffaloes with uterine torsion exhibited loss of appetite, constipation, colic, and straining. The incidence of torsion was mostly at first parity that accounting for 50% of cases. Regarding to the pregnancy stage, torsion mostly occurred at full term, where it primarily occurred in buffaloes at 3 years of age. Right-sided torsion ensued in 90% of all cases. The survival rate of the pregnant buffaloes was 90% (45 buffaloes), and 10% (5 buffaloes) was directed to the culling. Regarding to the fetal survival outcome, 30% of fetuses was survived where the other 70% was died. The hematological results denoted leucocytosis in Ut and Utc groups that attributed to neutrophilia, monocytosis, and eosinophilia. Biochemical assessments were denoted an augments in the levels of serum AST, LDH, CPK, creatinine, blood urea nitrogen, progesterone, cortisol, MDA, NO, TAC, blood lactate (bLac), TNF-α, and IL-6. Where there were a non significant fluctuations in the ALT, GGT, calcium, and phosphorus levels in the torsion cases. The significant declines in 17-ß-oestradiol, total proteins, albumin, magnesium, and GSH levels in the dams suffering from Ut were observed. The robust elevations in AST (156 ± 3.47) and bLac (16.12 ± 0.18) with severe hypomagnesaemia (2.69 ± 0.15) were evident in 10% of the torsion cases that directed to the culling. This can conclude that robust elevations in AST and bLac with severe hypomagnesaemia are factors denoting a poor prognosis for uterine torsion in the buffaloes.

Effects of intra-uterine infusion of proteolytic enzymes on selected cytokine concentrations, uterine inflammation, and fertility in postpartum water buffalo cows with subclinical endometritis.

The study was conducted to evaluate effects of intrauterine administration of proteolytic enzymes on endometrial inflammation and reproductive performance in postpartum water buffalo cows with subclinical endometritis (SCE). Cows (n = 38) with SCE (≥ 18 % PMN i.e.; polymorphonuclear cells) on day 21 postpartum (21 dpp), were allocated into treatment (TR; n = 19; intrauterine infusion of trypsin, chymotrypsin and papain in 20 ml normal saline on 21 dpp) and control (PC; n = 19; intrauterine administration 20 ml saline) groups. Cows without SCE (< 18 % PMN) were not treated and served as the negative control (NC; n = 30). Ultrasonography and sampling (endometrial cytology, uterine flushing, blood) were conducted on day 21 (before treatment) and 28 postpartum (28 dpp). The PMN % and uterine horn diameter were less on 28 dpp (compared with 21 dpp) in NC and TR group only. Cows with SCE had greater uterine concentrations of interleukin (IL)-1ß, IL-8 and tumor necrosis factor (TNF)-α; but lesser IL-10 than NC group on 21 and 28 dpp. There were greater serum IL-1ß and TNF-α concentrations on 28 dpp in cows with SCE than NC group. Uterine concentrations of IL-1ß were less, whereas IL-6 was greater following enzymatic treatment. Proteolytic enzyme treatment did not result in improvement in pregnancy rate compared with the PC group; however, days to conception were less in TR compared with the other two groups. In conclusion, results indicated a reduction in endometrial inflammation and days nonpregnant after proteolytic enzyme treatment in buffalo cows with SCE.

Prevalence of subclinical uterine pathologies diagnosed by biopsy and cytological and bacteriological findings in cyclic bitches.

The study was performed on 45 bitches in different cycle phases that were divided into the following groups: anoestrus (I, n=15), heat (pro-oestrus (n=7) or estrous (n=8) (II, n=15) and metestrus (III, n=15). Moreover, all experimental dogs were grouped according to their age: younger than 5 years (Y, n=35) and older than 5 years (O, n=10). The endometrial status was evaluated using cytological, bacteriological and biopsy samples obtained after ovariohysterectomy. The main uterine pathology diagnosed by biopsy was endometritis, since 40%-66% of bitches, independent of the experimental group, developed this condition. No significant differences were found among the cycle phase groups (p>0.05). By contrast, significant differences were found in the age groups; the prevalence of this pathology was higher in older bitches (p=0.0019). The general prevalence of cystic endometrial hyperplasia (CEH) and a normal endometrium (NE) was lower (6.7-26.7% vs 26.7-53.3%) in all groups, and no statistically significant differences were found between certain groups (p>0.05). The percentage of polymorphonuclear neutrophilic leukocytes (PMNs) in endometrial cytology was generally low (⟨ 2%) and did not differ significantly among the experimental groups (p=0.142). In general, a low degree of correlation was found between the diagnostic results by endometrial cytology and biopsy (Kappa Coefficient= 0.046). Positive bacteriological findings were found in approximately 50% of the bitches, independent of the cycle phase and health status of the endometrium. No correlation was found between the bacteriological and histopathological findings (p=0.883). In conclusion, uterine cytology is not a reliable diagnostic method to detect the subclinical inflammatory and degenerative uterine pathologies in cyclic bitches.

Participation of acetylcholine and its receptors in the contractility of inflamed porcine uterus.

This study analyzed the effect of inflammation on acetylcholine (ACh)-induced muscarinic receptors (MR)2 and MR3 conducted contractility of the porcine uterus. On Day 3 of the estrous cycle, either E.coli suspension (E.coli group) or saline (SAL group) was injected into uterine horns or laparotomy was performed (CON group). Eight days later, infected gilts developed severe acute endometritis. Compared to the period before ACh treatment, ACh (10-5 M) increased the tension in myometrium (MYO) and endometrium/myometrium (ENDO/MYO) of the CON group (P < 0.01) and in ENDO/MYO of the SAL group (P < 0.01), the amplitude in strips of the CON (P < 0.05) and SAL (MYO: P < 0.05, ENDO/MYO: P < 0.001) groups and the frequency in strips of the CON (MYO: P < 0.01, ENDO/MYO: P < 0.001) and SAL (P < 0.01) groups. In the E.coli group, ACh (10-5 M) reduced the amplitude in MYO (P < 0.05) and ENDO/MYO (P < 0.001), increased the frequency in MYO (P < 0.01) and ENDO/MYO (P < 0.001) and did not change (P > 0.05) the tension. ACh (10-5 M) in ENDO/MYO of the E.coli group, reduced the tension compared to the CON group (P < 0.05) and the amplitude compared to other groups (P < 0.001), while increased the frequency in relation to the SAL group (P < 0.05). MR2 antagonist (AF-DX 44 116) and ACh (10-5 M) reduced (by 16.92%, P < 0.01) the tension in MYO of the CON group and increased (P < 0.01) it in the E.coli group compared to the period before antagonist and ACh addition. In MYO of the SAL group, the tension was increased (P < 0.01) in response to MR3 antagonist (4-DAMP) and ACh (10-7, 10-6 M). In the E.coli group, these substances did not change (P > 0.05) the tension, but it was lower (P < 0.001) in MYO (ACh: 10-7 M) and ENDO/MYO (ACh: 10-5 M) than in the SAL group. MR2 or MR3 antagonists and ACh (10-5 M) increased (P < 0.05-0.001) the amplitude in strips of the CON and SAL groups and reduced it in the E.coli group (P < 0.001) compared to the period before antagonists and ACh use. This parameter in the E.coli group was lower (P < 0.001) after using MR2 or MR3 antagonists and ACh (10-6, 10-5 M) than in other groups. Both antagonists and ACh (10-5 M) reduced the frequency in the CON, SAL (P < 0.05) and E.coli (MR2 antagonist: P < 0.01, MR3 antagonist: P < 0.05) groups compared to period before antagonists and ACh addition. Data show that ACh reduces the contractility of the inflamed porcine uterus by MR2 and MR3, which suggests that pharmacological modulation of these receptors can be used to raise the contractility of an inflamed uterus.

Uterine responses and equine chorionic gonadotropin concentrations after two intrauterine infusions with kerosene post early fetal loss in mares.

Pregnancy loss during the normal lifespan of endometrial cups (∼37-120-150 days of gestation) may affect a mare's ability to conceive again in the same breeding season, as equine chorionic gonadotropin (eCG) secretion by retained endometrial cups can lead to abnormal ovulations and follicular growth. While intrauterine kerosene infusion has anecdotally been proposed as a treatment for endometrial cup retention, there are no controlled studies evaluating kerosene's ability to enhance endometrial cup regression following abortion. The objectives of this study were to assess uterine response, systemic side effects, and efficacy of intrauterine kerosene infusions after abortion. We hypothesized that kerosene infusions would hasten regression of endometrial cups without detrimental effects on the endometrium and the mare's general health. Twelve light-breed mares were enrolled in the study after an experimentally induced abortion with cloprostenol (n = 12) by 60 ± 2 days of gestation. Mares were randomly allocated to receive an intrauterine infusion with 500 mL of kerosene (n = 6) or 500 mL saline (n = 6) on days 21 and 35 after pregnancy termination. Uterine biopsies were collected at days 7, 21, 35, and 49 post-abortion to evaluate the degree of endometrial fibrosis with Picrosirius Red Stain and to be graded according to the Kenney & Doig 1986 classification. Furthermore, histomorphometry analysis of the endometrium lining, glandular epithelium and glandular density was performed. Endometrial lymphocyte B CD20+, lymphocyte T CD3+, and macrophage IBA-1+ cell populations were characterized by immunohistochemistry. Physical examinations, blood cell counts, and serum biochemistry were performed before, and for 2 days after each uterine infusion. Serum samples were collected for assessment of eCG concentrations. Continuous data were analyzed with MIXED procedure with repeated measures in SAS, categorical data with LOGISTIC procedure of SAS. Significance was set at p < 0.05. Kerosene infusion did not affect complete blood cell counts, serum chemistry parameters, or physical examinations. Concentrations of eCG decreased over time (p < 0.001), but there were no differences between groups or time by group interactions (p = 0.72). Histological evaluation of the uterus showed no signs of increased fibrosis or degeneration in the treatment group. In conclusion, while kerosene infusions did not appear to have detrimental effects on mare health, our findings suggest that the use of kerosene in the uterus does not enhance the regression of endometrial cups by 49 days post-abortion.

Ultrasonographic and hormonal characterization of reproductive health and disease in wild, semiwild, and aquarium-housed southern stingrays (Hypanus americanus).

OBJECTIVE: To characterize physical examination, plasma biochemical, and ultrasonographic findings in aquarium-housed, managed semiwild, and wild southern stingrays (Hypanus americanus) with and without reproductive disease. ANIMALS: Southern stingrays from aquarium (n = 48), lagoon (managed semiwild; 34), and wild (12) habitats. PROCEDURES: Limited, opportunistic prosections were performed of presumed anatomically normal wild southern stingrays and compared with findings for aquarium-housed stingrays with reproductive disease. Ultrasonographic video data from both groups were used to assign a score (1 to 5) indicating increasing severity of ovarian and uterine reproductive disease. Plasma total 17ß-estradiol, estrone, progesterone, and testosterone concentrations were measured with enzyme immunoassays validated for use in southern stingrays. RESULTS: Ultrasonographic ovarian scores were significantly correlated with uterine scores. No reproductive disease was detected in semiwild or wild stingrays, but 65% (31/48) of aquarium-housed stingrays had developing or advanced reproductive disease (ie, ultrasonographic ovarian or uterine score of 4 or 5). Significant correlations were identified between ovarian and uterine disease status and plasma concentrations of all steroid hormones except testosterone. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggested that ultrasonography and plasma hormone concentrations may be useful in the identification of reproductive disease and determination of disease severity in southern stingrays.

Effects of pentoxifylline supplementation to semen extender on post-breeding inflammation response assessed by endometrial cytology and vascular perfusion in mares.

The aim was to evaluate effects of addition of pentoxifylline to skimmed milk semen extender on uterine inflammatory response. Thirty-six estrous cycles of 15 mares were randomly divided into five groups for artificial insemination (AI): Control: mimicking the AI procedure (n = 7); Extender: deposition of skimmed milk based extender (n = 7); Extender + PTX: skimmed milk based extender plus pentoxifylline (7.18 mM; n = 8); Semen: semen diluted with extender without pentoxifylline (n = 7), and Semen + PTX: semen diluted with extender containing pentoxifylline (n = 7). Mares in estrus were examined by trans-rectal palpation and using ultrasonography, and ovulation was induced. Uterine hemodynamics were assessed immediately before ovulation induction (T-30), immediately before AI (T0), 2 (T2), 6 (T6), 12 (T12), 24 (T24) and 48 (T48) h after AI. Endometrial samples were collected 6 h after AI, and slides were stained and examined to determine percentage of PMN. Pentoxifylline had no additional effect on vascular perfusion. There was a major inflammatory response with pentoxifylline treatment that was greater than that of the control group. In the group treated with Extender + PTX, there were more PMN (57.98 ±â€¯9.42%) than in the group treated with Extender (20.20 ±â€¯6.63%) and in the Semen + PTX group more PMN (82.84 ±â€¯5.71%) than in the Semen-treated group (47.83 ±â€¯10.61%). These findings indicate the addition of pentoxifylline does not stimulate blood flow; however, it induces a greater immune defense response because more neutrophils migrate to the uterine lumen.

Endometrial histopathology, bacteriology and cytology outcomes in mares with early embryonic death (EED): a field study.

Early embryonic death (EED) is one of the causes of infertility in the mare. We compared endometrial environment in 9 mares with EED and 13 mares in diestrus phase. Cotton swab (CS), cytobrush (CB) and uterine biopsy (B) samples were obtained for the cytological, bacteriological and histopathological examinations. In the first step we compared CS and CB methods to biopsy as a reference method, as B revealed the highest number of positive results in cytological and bacteriological examinations in both groups. In turn, we also compared cytological, bacteriological and histopathological findings between EED and control animals using the B sampling. Although the differences between these groups were not statistically significant (p≥0.05), there was a tendency to a higher prevalence of subclinical endometritis in the control group, than in the EED group (62% vs 22%). In general, positive bacteriological results were similar in both groups (62% vs 55%), whereas positive cytological results were higher in the control group (62% vs 22%; p≥0.05). In histopathological examination in EED mares endometrial degeneration was better expressed (all mares were with grades IIB and III on the Kenney-Doig scale); however, the differences between both groups were not statistically significant (p≥0.05). We could not confirm a clear difference in uterine environment between the two groups. Moreover, the uterine biopsy seemed to be the most reasonable sampling method for diagnosis of endometrial state.

Assessment of the optimal age for a preventive ultrasonographic screening of the uterine health in bitches.

Studies about prevalence of uterine pathologies in bitches are scarce. Although correlation between age and uterine disorders was documented, the most suitable age for a preventive sonographic screening has not been proposed yet. Present study aimed to estimate the eligible age for an ultrasonographic screening of uterine abnormalities in dogs. Data regarding ultrasound examination and clinical records of non-pregnant intact females were retrospectively analysed. The age of each bitch was expressed as age ratio (actual/maximum age expected for the respective breed). The cut-off age ratio was determined by a ROC curve for overall uterine abnormalities. Frequencies of different abnormalities below and over the cut-off derived from the ROC curve were calculated and statistically analysed by chi-Square and OR. Prevalence of three categories of ultrasonographic findings was as follows: cystic endometrial hyperplasia (CEH) 18%; uterine collections (UC) 10.5%; masses (M) 1.3%. By the cut-off age ratio (0.325), derived by ROC curve (AUC = 0.91; SP 84.23%; SE 79.2%; PPV 83.4%; NPV 80%), 228 cases were divided into two subgroups: bitches over (exposed group: n.83) and below cut-off (control group: n.145). All abnormalities resulted more frequent in exposed group: OR was 24.96 (p < 0.0001: 71.1% over vs. 9% below cut-off) for overall abnormalities; 13.68 (p < 0.0001: 40.9% vs. 4.8%) for CEH; 6.13 (p < 0.002: 21.7% vs. 4.1%) for UC; 12.65 (p = 0.09: 3.6% vs. 0%) for M. Cystic endometrial hyperplasia represents the most common finding in adult bitches, followed by UC. A preventive sonographic screening for uterine abnormalities should start from 33% of expected longevity to preventively select animals requiring further evaluations.