Histological Changes in the Uterus during Postpartum in the Mare

An histological study of the postpartum period in 29 mares was carried out. Uterine biopsies were taken daily during the first 10 days postpartum in a total of 87 samples. At day 0, equine endometrium was characterized in the surface by the presence of regularly ordered microcaruncles; the stratum spongiosum was oedematous and contained distended and scarce glands. Degenerative changes in microcaruncles and endometrial glands were present on day 1 postpartum. The epithelium of the microcaruncles from 2 to 5 days postpartum showed cytoplasmic vacuolization, karyorrhexis and an inflammatory reaction with neutrophils and phagocitic cells. On day 7 postpartum, the histology of the endometrium was similar to the normal proestrus with cuboidal luminal epithelium and an oedematous stromal tissue. The changes are usually completed within days 9 and 10 postpartum with the histologically typical appearance of estrus in mares.     

Effects of Neodymium:Yttnum Aluminum Garnet Laser Irradiation on Endometrium and on Endometrial Cysts in Six Mares

Effects of neodymium:yttrium aluminum garnet (Nd:YAG) laser irradiation on equine endometrium were evaluated In vitro and in six mares with endometrial cysts. The Nd:YAG laser was applied to six endometrial sites, in each of five uterine specimens, with power densities of 5659 to 33,954 J/cm². Depth of tissue ablation was measured and graded on histologic sections of the tissue lesions. Power density had a significant effect on the depth of tissue ablation (p < .001). Grade 3 lesions (full-thickness ablation of the endometrium) were created with energy densities of 16,977 to 33,954 J/cm². Six mares had endometrial cysts treated by photoablation. Two of the four mares that were reproductively sound but barren, despite appropriate breeding, produced foals after treatment. One mare remained reproductively unsound after treatment, and another mare that was treated postpartum was bred successfully.     

Expression of Aquaporin Water Channels in Equine Endometrium is Differentially Regulated During the Oestrous Cycle and Early Pregnancy

The expression of 12 different aquaporin subtypes in equine endometrium was examined at the mRNA and protein level. Endometrial samples were obtained during anoestrus, oestrus, 8, and 14 days after ovulation in non‐pregnant mares, and 14 days after ovulation in pregnant mares. Quantitative PCR revealed a time‐dependent pattern for all aquaporin subtypes examined except for AQP10 and 12. AQP3, 5 and 7 showed highest mRNA abundance 8 days after ovulation, while AQP0 and 2 were most abundant at Day 14 of the cycle in non‐pregnant mares. At 14 days of pregnancy, AQP1, 4, 8, 9 and 11 displayed highest expression levels. Western blot analysis confirmed protein expression of AQP0, 2 and 5. Immunohistochemistry localized protein expression to luminal and glandular epithelial and stromal cells. AQP0 staining intensity was highest in samples obtained on Day 14 of the oestrous cycle. AQP2 immunoreactivity seemed to be stronger in samples collected 14 days after ovulation from non‐pregnant animals, in particular luminal epithelial staining. Samples collected 8 days after ovulation from cyclic animals were characterized by intense AQP5 staining of glandular epithelium, predominantly in the deeper glands. Progesterone treatment of anoestrous mares did not enhance expression of AQPs, indicating that factors other than progesterone are required for the up‐regulation of certain AQP subtypes during dioestrus. In conclusion, it seems that an equine‐specific collaboration of aquaporin subtypes contributes to changes in endometrial fluid content occurring throughout the oestrous cycle and contributes to endometrial receptivity during early pregnancy in the mare.     

The expression of interferon-stimulated gene 15 in equine endometrium

Establishment of pregnancy is critically dependent upon a precisely orchestrated embryo-maternal interaction leading to a receptive uterine environment. The up-regulation of the interferon-stimulated protein 15 kDa (ISG15) during pregnancy has been described in various species and has been hypothesized to be part of the molecular repertoire that makes the uterus receptive to conceptus development. In the current study, the expression of ISG15 and enzymes involved in ISG15ylation was examined at the mRNA and protein level in equine endometrium at Day 14 of the luteal phase and at Day 14 and 50 of pregnancy. ISG15 mRNA showed a 2.63-fold higher expression at Day 14 of pregnancy when compared to Day 14 of the cycle, while mRNA abundance at Day 50 of pregnancy was unchanged compared to Day 14 of the cycle. Upon Western blot analysis using anti-ISG15 antibody, several higher molecular weight bands could be observed, representing proteins conjugated to ISG15. No free ISG15 could be detected. The pattern of ISG15 reactive proteins differed from those observed in non-uterine samples. Upon immunohistochemistry, ISG15 reactive proteins located primarily to luminal and glandular epithelial cells, while stromal cells showed weaker staining. In conclusion, the expression of ISG15-conjugated proteins in equine endometrium did not differ between cyclic and pregnant 14 days after ovulation and Day 50 of pregnancy. It is hypothesized that the unique subset of ISG15ylated proteins expressed in endometrial tissue contributes to normal cellular function and that, unlike other species, the modification of ISG15-conjugated proteins is not an active contributor to conceptus-maternal interaction in the mare.     

Study of the duration and distribution of equine influenza virus subtype 2 (H3N8) antigens in experimentally infected ponies in vivo.

The purpose of this experiment was to study the duration and distribution of equine influenza virus in actively infected ponies over a 3 wk period. Pony foals (6-8 mo old) were infected experimentally by nebulizing equine influenza subtype-2 virus ultrasonically through a face mask. Successful infection was clinically apparent as each of the foals (n = 6) had a febrile response, a deep hacking cough and mucopurulent nasal discharge for 7 to 10 d. The virus was isolated from nasopharyngeal swabs of all the ponies 3 and 5 d after infection and all the ponies seroconverted to the virus. Samples were taken from the nasopharynx, mid-trachea and the mainstem bronchus with cytology brushes through an endoscope as well as from bronchoalveolar lavage fluid. On days 3 to 7 post-infection, ciliacytophtorea (the presence of cilia and ciliated plates separated from columnar epithelial cells) was recognized on routine cytological stain. Indirect immunoperoxidase staining utilizing polyclonal antibodies demonstrated viral antigen in intact and fragmented ciliated epithelial cells and in fragments of ciliated plates. The infected cells and cell fragments were particularly evident on days 3 and 5 post-infection in the nasopharynx, mid-trachea and mainstem bronchus and on days 3 to 7 post-infection in the bronchoalveolar lavage samples. On days 7 and 21 post-infection, viral antigen was identified in vacuoles of alveolar macrophage-like cells collected by bronchoalveolar lavage. It can be concluded from this study that equine influenza virus can infect not only the upper airways but also the bronchial epithelium and that viral antigen can persist up to 21 d post-infection.     

The uterus and early pregnancy failure in the mare

The environment the equine conceptus finds itself in when it arrives in the uterus some 6 days after ovulation will determine if it will thrive or die. The uterus will be its home for approximately the next 11 months and as such it needs to both coordinate the growth of the placenta and ensure there is enough nourishment passing to this organ for the fetus to develop normally. The trophoblast, endometrium, maternal ovaries and, later in pregnancy, the fetal gonads, all play roles in the hormonal changes that orchestrate these events. Although failures of these processes later in pregnancy can have catastrophic effects for the fetus, it is in early gestation that the foundations for a successful pregnancy are laid. This paper therefore concentrates on some of the noninfectious influences the uterus may have on survival of the young conceptus.     

Endometrial Phospholipase A2 Activity During the Oestrous Cycle and Early Pregnancy in Mares

The aim of this study was to determine phospholipase A2 (PLA2) kinetics and activity in the mare’s endometrium during the oestrous cycle and early pregnancy. Phospholipase A2 is responsible for the liberation of arachidonic acid from phospholipids, which is the first limiting step in prostaglandins synthesis. Phospholipase A2 activity was measured using an assay based on the liberation of oleic acid from 1‐palmitoyl‐2‐[¹⁴C] oleoyl phosphatidylcholine. The enzyme was shown to be calcium dependent, to have an optimum pH of 8 and an apparent Michaelis constant of 127 μm . Enzyme activity was low in the endometrium of early luteal phase tissue but increased significantly (p < 0.001) during the late luteal phase (5.39 ± 0.16; 3.48 ± 0.33, 6.85 ± 0.59, and 9.96 thinsp;± thinsp;1.23 thinsp;nmol oleic acid released/mg protein at oestrus, and Days 3, 8 and 14 after ovulation, respectively). The mean PLA2 activity in endometrial tissue from pregnant mares (4.23 ± 0.74) was significantly lower (p < 0.01) than from cyclic animals during late dioestrus (9.96 ± 1.23). The results indicate that PLA2 activity in equine endometrium changes with the stage of the oestrous cycle and thus may be influenced by systemic hormone concentrations. The inhibitory effects of conceptus products on secretion of prostaglandin during early pregnancy were associated with a competitive inhibitor that decreased endometrial PLA2 activity.     

Determination of intestinal viability

Operations for intestinal ischemia are frequently done by veterinarians. In equine surgery those conditions commonly producing ischemia are intussusception, volvulus, bowel obstructions, and incarcerated hernias. In an attempt to predict intraoperative bowel viability after the restoration of circulation, a variety of adjuvant methods have been investigated. There is little question that of the techniques currently available, sodium fluorescein injected intravenously approaches the ideal in predicting nonviability in humans and in most animal models. Furthermore, it is safe, takes little operating time, is inexpensive, and is ubiquitously available; in addition, the only special equipment needed is a long-wave ultraviolet lamp. Reliance on this method would seldom result in nonviable bowel being left in situ. Furthermore, the use of fluorescein would minimize the unnecessary resection of viable intestine and, thereby, minimize the postoperative nutritional problems that may occur, especially in equines. The report of Sullins et al questions the accuracy of the fluorescein technique in predicting viability and nonviability in horses and also raises the question as to whether the prediction of viability, after the restoration of intestinal blood flow, also predicts normal intestinal function. Further investigations will be needed to confirm or refute these observations.     

Equine placenta – A clinician's perspective. Part 1: Normal placenta – Physiology and evaluation

A complex organ, the equine placenta is responsible for fetal nourishment, protection from external and internal insults, and the production and/or metabolisation of various hormones. The endometrial cups are unique structures of the equine placenta that are responsible for producing an essential hormone for equine pregnancy, equine chorionic gonadotropin. Since mares have epitheliochorial placentae, with 6 layers of tissue separating maternal from fetal circulation, almost the entire surface of the chorioallantois must be attached to the maternal endometrium in order to support a developing fetus adequately. The only avillous areas of the normal chorioallantois are: the cervical star, sites of the endometrial cups, areas facing oviductal papillae, folds overlying major allantoic vessels, and fetal foot PADs (placental areas of degeneration). There are characteristic differences between the gravid and the nongravid horn of the chorioallantois. Allantoic vesicles, allantoic pouches, hippomanes, amniotic plaques and yolk sac remnants are normal features of the equine placenta. The clinician should thoroughly examine the entire placenta immediately after its expulsion. The most important aspect of this evaluation is to check for completeness of the chorioallantois, along with identifying any pathological lesions on the fetal membranes or the umbilical cord.     

Administration of hCG 5 days after Breeding and Reproductive Performance in Nulliparous Dairy Goats

The objective of this study was to investigate the effect of hCG administration 5 days after breeding on plasma progesterone (P4) concentration and reproductive performance of oestrous-induced nulliparous dairy goats. A total of 59 nulliparous goats (36 Alpine and 23 Saanen) received intravaginal sponges with 60 mg medroxyprogesterone acetate for 9 days plus 200 IU equine chorionic gonadotrophin (eCG) and 22.5 microg d-cloprostenol 24 h before sponge removal. After detection of oestrus (day of oestrus = day 0) and breeding, 49 females were randomly assigned, according to the breed, into two treatments (T1 and T2). In T1 (n = 25) and T2 (n = 24), animals received intramuscular injection of 1 ml of saline solution (control) or 250 IU hCG, respectively, 5 days after breeding. Plasma P4 concentration (ng/ml) was determined from blood sampled on days 0, 5, 7, 13, 17, 21, 28 and 45 after breeding. Animals were scanned by transrectal ultrasound (5 MHz probe) on days 35 and 70 after breeding for detection of pregnancy. Plasma P4 concentration did not differ (p > 0.05) between treatments in all days, but it was increased (p < 0.05) in Saanen than in Alpine goats from days 13 to 45. Pregnancy and parturition rates, litter size and gestation period were similar (p > 0.05) to treatments and breeds. Results of this study indicate that human chorionic gonadotrophin (hCG) administration 5 days after breeding did not significantly alter reproductive performance in dairy nulliparous goats and that plasma P4 differed between Saanen and Alpine goats.     

Absorption of bovine colostral immunoglobulins G and M in newborn foals

The uptake of colostral IgG and IgM, their serum half-lives, and the rates of endogenous synthesis of IgG and IgM were evaluated in 6 newborn foals fed bovine colostrum (principals) and 6 foals allowed to suckle their dams (controls). The principal foals were fed 400 ml of bovine colostrum (IgG, 10,000 mg/dl and IgM, 200 mg/dl) at 2-hour intervals, from 2 to 20 hours after foaling (total dose, 4 L). Serum IgG and IgM concentrations were determined by single radial immunodiffusion from birth to 98 days of age. At foaling, principal foals had no detectable serum equine IgG, but 1 control foal had serum equine IgG of 185 mg/dl. After ingestion of colostrum, there was no significant difference in the maximal serum bovine IgG concentration (range, 1,350 to 3,300 mg/dl) in the principal foals, and maximal serum equine IgG concentration in the control foals (range, 500 to 6,000 mg/dl). The calculated biological bovine and equine IgG half-life in the principal and control groups was 9.4 and 26 days, respectively. Endogenous IgG synthesis was first detected in 1 principal foal at 3 days of age, but was detected first between 28 and 42 days in the other principal foals. Starting on day 56 there was no significant difference in serum equine IgG concentration between groups. At foaling, foals in both groups had low equine IgM concentrations. In the control foals, there was marked individual variation in the increases in equine IgM concentration (range, 5 to 73 mg/dl) after ingestion of colostrum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation of equine coital exanthema virus (equine herpesvirus 3) from the nostril of a foal

The virus causing equine coital exanthema (equine herpesvirus 3) was isolated from a lesion on the nostril of a 2-month-old foal. One week after the mare had returned from a stallion station, vesicular lesions developed on her vulva. They were diagnosed clinically as coital exanthema, and 5 days later a lesion developed on the nostril of her foal. This case is an example of horse-to-horse transmission of coital exanthema virus without coitus. A laboratory diagnosis is necessary to differentiate viruses that cause vesicular lesions about the oral and nasal cavities of horses.     

Histology of the endometrium in cows in normal and pathologic puerperium

There were repeated histological bioptic examinations of the endometrium during the period from the second to twentieth day post partum (p.p.) in cows with afterbirth retention (n = 5), with developing puerperal endometritis (n = 5), and with a physiological course of puerperium (n = 5). In cows with a physiological course of puerperium, a finished process of epithelization of endometrium with a morphological and functional consolidation of the epithelium could be observed on the 20th day p.p. In cows with disturbed puerperium, the surface epithelium still exhibited signs of disintegration and a low secretion activity. The endometria of both groups of cows with pathological puerperium had a high degree of infiltration with neutrophil granulocytes: in cows with retained afterbirth this picture was characteristic already the second day p.p. On the other hand, the endometrium of cows with physiological puerperium was characterized by a higher occurrence of folliculoid nodules as early as on the tenth day p.p., and a rapid increase in the number of fat cells and activated histiocytes--macrophages could be observed in the endometrium of these cows between the 15th and 20th day p.p. The dynamics of occurrence of mobile cells in the studied groups of cows confirmed their importance for a physiological course of the restitution of endometrium during the puerperium of cows.     

Matrix Metalloproteinase (MMP)-2 and MMP-9 Activity in the Canine Uterus Before and During Placentation

The aim of the present study was to demonstrate the presence and localization of MMP-2 and -9 by means of RT-PCR and immunohistochemistry (IHC) within the canine uterus from the pre-implantation stage until mid-gestation and to determine MMP-2 and -9 activities by means of zymography. For this purpose, samples of the uterus and salpinx from bitches were obtained after ovariohysterectomy. Pre-implantation stages (5-12 days after mating, n = 11) were determined by verifying embryos after flushing the uterus. Further groups were determined as implantation (15-19 days after mating, n = 9), post-implantation (20-30 days after mating, n = 9) and placental stages (30-45 days after mating, n = 3). A non-pregnant group (17-30 days after mating, n = 4) served as control. MMP-2 and -9 positive cells were detected in all specimens from pregnant and nonpregnant bitches, however, with different distributions. MMP-2 was present in endothelium and smooth muscles of blood vessels and the myometrium of pregnant and nonpregnant bitches, additionally in the surface epithelium of the oviduct. The latter also stained positive for MMP-9. During placentation, MMP-2 was detected mainly in fetal blood vessels and trophoblastic cells. Higher MMP-2 activity was observed in the endometrium and myometrium of all pregnant groups compared with the nonpregnant group (p < 0.05). The pregnant groups did not differ significantly from each other (p > 0.05). MMP-9 was present in blood vessels, smooth muscle cells and epithelia, such as maternal surface epithelial cells, uterine crypts and glands. During placentation, the deep uterine glands and the epithelium of the glandular chambers were immunoreactive to MMP-9. Highest MMP-9 activities were reached in the endometrium of the pre-implantation group (23.2% of total MMP-9) and placental parts (33.3%).     

Equine placenta – A clinician's perspective. Part 2: Abnormalities

Any insufficiency of the equine placenta has dramatic effects on the developing equine fetus. Placental abnormalities, such as the separation of the chorioallantois from the maternal endometrium or torsion of the umbilical cord, lead to fetal demise, premature labour or abortion. These conditions are each associated with characteristic lesions on the equine placenta, which can be found during a detailed examination. These findings can be very helpful for diagnosing problems and implementing appropriate treatments for mares and affected newborn foals. Furthermore, the retention of the entire placenta or any small fragment thereof can cause metritis, laminitis and sepsis. The prompt diagnosis and aggressive treatment of this condition is necessary to save the mare from becoming seriously ill. Therefore, a thorough evaluation of the equine placenta is a crucial element of the post partum evaluation of every brood mare.     

Passage of equine herpesvirus-1 in suckling mouse brain enhances extraneural virus growth and subsequent hematogenous neuroinvasion

Intracerebral inoculation of field-isolates as well as established strains of equine herpesvirus-1 (EHV-1) in suckling mice results in viral replication in neurons and glial cells and induces encephalitis. By intraperitoneal (i.p.) inoculation, no histological lesion was observed in the central nervous system (CNS) in suckling mice with the EHV-1 HH1 strain (HH1), whereas a neuroadapted variant (NHH1) produced by serial passage of HH1 in the mouse brain caused severe encephalomyelitis after i.p. inoculation. The purpose of this study was to determine the route of neuroinvasion after i.p. inoculation of NHH1 and to clarify the effects of the brain passage on viral neuroinvasion. NHH1, but not HH1, targeted splenic and pulmonary macrophages and omental fat cells on days 1 and 2 post-inoculation (p.i.). From days 1 to 3 p.i., cell-associated viremia was occurred in NHH1-infected mice, but not in HH1-infected mice. On day 4 p.i., viral antigen was detected in a few endothelial cells, perivascular glial cells and neurons in the CNS in NHH1-infected mice. The number of viral antigen-positive cells increased markedly after day 5 p.i. In contrast, no viral antigen-positive cell was detected in the CNS in HH1-infected mice, except for a few nerve cells in the thoracic cord on day 4 p.i. These results suggest that NHH1 neuroinvasion is hematogenous and is correlated with enhanced extraneural virus growth.     

产后奶牛子宫内膜的扫描电镜观察

应用扫描电镜观察了 5头奶牛产后不同时期子宫内膜的变化。结果发现 ,分娩后早期 ,奶牛子宫内膜部分破损 ,与破损部位相邻的细胞在产后 1周内相继脱落 ;未破损内膜上皮细胞表面的纤毛和微绒毛形状不整、数量减少。产后2周 ,基底细胞开始生长 ,至产后 4 5 d时生长完成 ;产后 4周 ,内膜破损区域缩小 ,上皮细胞表面纤毛和微绒毛形状规则、数量增加 ;产后 4 5 d,偶见上皮细胞缺失 ;至产后 6 0 d,子宫内膜上皮细胞完整、排列有序、被覆纤毛和微绒毛。上述结果表明 ,奶牛子宫内膜在分娩后 6 0 d内完成修复。     

Effect of Weaning to Oestrus Interval and Equine Chorionic Gonadotropin on Vaginal Electrical Impedance During Peri‐oestrus in Sows

The influence of weaning to oestrus interval, its interaction with parity and equine chorionic gonadotropin (eCG) on changes of vaginal impedance in sows after weaning was examined. The impedance measurements were carried out by a four‐terminal method. Sows were monitored for oestrus via exposure to a sexually mature boar. The interval from weaning to oestrus was longer in primiparous than multiparous sows (p < 0.01). A significant negative correlation was found between the interval from weaning to oestrus and parity. Repeated measures analysis showed that the interval from weaning to oestrus and parity and their interactions had a significant effect on the vaginal impedance in peri‐oestrus. The vaginal impedance during pro‐oestrus gradually decreased in all groups of sows with the weaning to oestrus interval from 4 to 8 days (p < 0.05). In the subsequent period, the vaginal impedance increased and was significantly lower from 1 to 3 days after oestrus onset in sows with the weaning to oestrus interval 7–8 days than 4–6 days. Similarly, the vaginal impedance during pro‐oestrus gradually decreased in all groups of sows with parity 1–5 (p < 0.01). In the next period, the vaginal impedance increased and was significantly lower from 2–3 days after oestrus onset in sows of parity 1 than parity 2–5. Repeated measures analysis showed that eCG treatment had a significant effect on the vaginal impedance in peri‐oestrus. Sows treated with eCG displayed the decrease and increase of vaginal impedance due to oestrus onset earlier than untreated sows. The results indicate that the weaning to oestrus interval, its interaction with parity and eCG markedly affect the vaginal impedance in sows during peri‐oestrus.     

Morphologic stages of the equine embryo proper on days 17 to 40 after ovulation

OBJECTIVE: To describe the gross and histologic changes that develop in the equine embryo proper (ie, the portion of the embryo that becomes the fetus) from days 17 to 40 after ovulation and to compare the external features of equine embryos with those of porcine, ovine, and human embryos. SAMPLE POPULATION: 34 embryos collected from mixed-breed pony mares. PROCEDURE: External features for each embryo proper, including length, number of branchial arches, growth of appendages, face and head features, and body features, were examined, using a dissecting microscope, for embryos collected on days 17 to 40. Internal features were histologically examined by serially sectioning embryos collected on days 20 to 35. RESULTS: Number of embryos recovered for each day ranged from 1 to 5. The initial detection of features was not related closely to age; typically, the first attainment of a given body length or characteristic varied over a 3-day period among embryos. Similarly, the period during which individual characteristics for a given Carnegie stage were attained ranged from 3 to 6 days. Age at first appearance of a characteristic was greater for equine embryos than ages reported for ovine and porcine embryos but less than for human embryos. Indicators of age included number of pairs of branchial arches, all limb buds present, retinal pigmentation, and prominence of the pontine flexure. CONCLUSIONS: No embryologic structures or changes were found that could be considered unique to equine embryos on days 17 to 40 after ovulation.