The effect of select seminal plasma proteins on endometrial mRNA cytokine expression in mares susceptible to persistent mating‐induced endometritis

In the horse, breeding induces a transient endometrial inflammation. A subset of mares are unable to resolve this inflammation, and they are considered susceptible to persistent mating‐induced endometritis PMIE Select seminal plasma proteins cysteine‐rich secretory protein‐3 (CRISP‐3) and lactoferrin have been shown to affect the innate immune response to sperm in vitro. The objective of this study was to determine whether the addition of CRISP‐3 and lactoferrin at the time of insemination had an effect on the mRNA expression of endometrial cytokines in susceptible mares after breeding. Six mares classified as susceptible to PMIE were inseminated during four consecutive oestrous cycles with treatments in randomized order of: 1 mg/ml CRISP‐3, 150 μg/ml lactoferrin, seminal plasma (positive control) or lactated Ringer's solution (LRS; negative control) to a total volume of 10 ml combined with 1 × 10⁹ spermatozoa pooled from two stallions. Six hours after treatment, an endometrial biopsy was obtained for qPCR analysis of selected genes associated with inflammation (pro‐inflammatory cytokines interleukin (IL)‐1β, IL‐8, tumour necrosis factor (TNF)‐α, interferon (INF)‐γ, anti‐inflammatory cytokines IL‐1RN and IL‐10, and inflammatory‐modulating cytokine IL‐6). Seminal plasma treatment increased the mRNA expression of IL‐1β (p = .019) and IL‐8 (p = .0068), while suppressing the mRNA expression of TNF (p = .0013). Lactoferrin also suppressed the mRNA expression of TNF (p = .0013). In conclusion, exogenous lactoferrin may be considered as one modulator of the complex series of events resulting in the poorly regulated pro‐inflammatory response seen in susceptible mares.     

Ovulation,Pregnancy Rate and Early Embryonic Development in Vernal Transitional Mares Treated with Equine‐ or Porcine‐FSH

The objective of this study was to compare the efficacy of purified equine‐ and porcine‐FSH treatment regimes in mares in early vernal transition. Mares (n = 22) kept under ambient light were examined ultrasonographically per‐rectum, starting January 30th. They were assigned to one of two treatment groups using a sequential alternating treatment design when a follicle ≥ 25 mm was detected. In the eFSH group, mares were treated twice daily with equine‐FSH, and in the pFSH group mares were treated twice daily with porcine‐FSH; treatments were continued until follicle(s) ≥ 35 mm, and 24 h later hCG was administered. Oestrous mares were inseminated with fresh semen and examined for pregnancy on days 11–20 post‐ovulation. In the eFSH group, 11/11 (100%) mares developed follicle(s) ≥ 35 mm, 8/11 (73%) ovulated and 6/8 (75%) conceived. In the pFSH group, 5/11 (45%) developed follicle(s) ≥ 35 mm, 4/11 (36%) ovulated and 3/4 (75%) conceived. Treatment with eFSH resulted in a greater ovarian stimulation; higher number of pre‐ovulatory‐sized follicles, higher number of ovulations and higher number of embryos (p < 0.05). Following ovulation, serum progesterone concentrations were correlated with the number of CLs and supported early embryonic development; maternal recognition of pregnancy occurred in all pregnant mares. We concluded that eFSH can be used to effectively induce follicular growth and ovulation in vernal transitional mares; however, if bred, diagnosis and management of twins’ pregnancies would be required prior to day 16 because of the increased risk of multiple embryos per pregnancy. Conversely, the current pFSH treatment regime cannot be recommended.     

Transplacental Transfer of Iron in the Water Buffalo (Bubalus bubalis): Uteroferrin and Erythrophagocytosis

The objectives of this investigation were to understand transplacental transport of iron by secreted uteroferrin (UF) and haemophagous areas of water buffalo placenta and clarify the role(s) of blood extravasation at the placental‐maternal interface. Placentomes and interplacentomal region of 51 placentae at various stages of gestation were fixed, processed for light and transmission electron microscopy, histochemistry and immunohistochemistry. Haemophagous areas were present in placentomes collected between 4 and 10 months of pregnancy. Perl’s reaction for ferric iron was negative in placentomes, but positive in endometrial glands. Positive staining for UF indicated areas in which it was being taken up by phagocytosis and/or fluid phase pinocytosis in areolae of the interplacentomal mesenchyme, with little staining in endometrial stroma. Imunohistochemistry detected UF in trophectoderm of haemophagous regions of placentomes and in other parts of the foetal villous tree, but the strongest immunostaining was in the epithelial cells and lumen of uterine glands. Ultrastructural analyses indicated that erythrophagocytosis was occurring and that erythrocytes were present inside cells of the chorion that also contained endocytic vesicles and caveolae. Results of this study indicate that both the haemophagous areas of placentomes and the areolae at the interface between chorion and endometrial glands are important sites for iron transfer from mother to foetal‐placental tissues in buffalo throughout pregnancy.     

Endothelin receptor subtype A blockade does not affect the haemodynamic recovery from haemorrhage during xenon/remifentanil or isoflurane/remifentanil anaesthesia in dogs

ObjectiveTo test the compensatory role of endothelin-1 when acute blood loss is superimposed on anaesthesia, by characterizing the effect of systemic endothelin receptor subtype A (ET_A) blockade on the haemodynamic and hormonal responses to haemorrhage in dogs anaesthetized with xenon/remifentanil (X/R) or isoflurane/remifentanil (I/R).Study designProspective experimental randomized controlled study.AnimalsSix female Beagle dogs, 13.4 ± 1.3 kg.MethodsAnimals were anaesthetized with remifentanil 0.5 μg kg^?1 minute^?1 plus either 0.8% isoflurane (I/R) or 63% xenon (X/R), with and without (Control) the systemic intravenous endothelin receptor subtype A antagonist atrasentan (four groups, n = 6 each). After 60 minutes of baseline anaesthesia, the dogs were bled (20 mL kg^?1) over 5 minutes and hypovolemia was maintained for 1 hour. Continuous haemodynamic monitoring was performed via femoral and pulmonary artery catheters; vasoactive hormones were measured before and after haemorrhage.ResultsIn Controls, systemic vascular resistance (SVR), vasopressin and catecholamine plasma concentrations were higher with X/R than with I/R anaesthesia at pre-haemorrhage baseline. The peak increase after haemorrhage was higher during X/R than during I/R anaesthesia (SVR 7420 ± 867 versus 5423 ± 547 dyne seconds cm^?5; vasopressin 104 ± 23 versus 44 ± 6 pg mL^?1; epinephrine 2956 ± 310 versus 177 ± 99 pg mL^?1; norepinephrine 862 ± 117 versus 195 ± 33 pg mL^?1, p < 0.05). Haemorrhage reduced central venous pressure from 3 ± 1 to 1 ± 1 cmH₂O (I/R, ns) and from 8 ± 1 to 5 ± 1 cmH₂O (X/R, p < 0.05), but did not reduce mean arterial pressure, nor cardiac output. Atrasentan did not alter the haemodynamic and hormonal response to haemorrhage during either anaesthetic protocol.Conclusions and clinical relevanceSelective ET_A receptor blockade with atrasentan did not impair the haemodynamic and hormonal compensation of acute haemorrhage during X/R or I/R anaesthesia in dogs.     

Ontogeny of hepatic bovine growth hormone receptors in cattle

A series of studies examined the binding characteristics and ontogeny of hepatic growth hormone binding sites in dairy bulls on d 2, 30, 180, and 365 of age. Binding of iodinated recombinant bovine growth hormone ([125I]rbGH) to liver membrane receptors was membrane protein-dependent. Receptors were considered growth hormone-specific, because physiological concentrations of bovine prolactin (bPRL) failed to displace [125I]rbGH from bovine hepatocyte membranes. Only 50% of [125I]rbGH was bound reversibly to hepatic microsomes. Addition of dithiothreitol (DTT) to the receptor-assay buffer increased the binding of [125I]rbGH to hepatic membranes in a time-dependent manner. Moderate concentrations of Ca++ and Mg++ in the receptor-assay buffer had no detectable effects on binding of [125I]rbGH to hepatic microsomes. In growing dairy bulls, specific binding of [125I]rbGH per milligram of membrane protein increased from 1.9 +/- 1.8% at d 2 to 14.1 +/- 1.8% at d 180 and then declined to 5.2 +/- 1.6% at d 365. Likewise, concentration of insulin-like growth factor (IGF)-I in serum was low during the 1st mo of age (d 2, 13.3 +/- 8.8 ng/ml; d 30, 9.7 +/- 8.8 ng/ml), but it became maximal at d 180 (151.0 +/- 8.8 ng/ml). Circulating concentrations of IGF-II increased linearly during the 1st yr of growth. Serum concentrations of GH, triiodothyronine, and thyroxine declined from 39.9 +/- 6.5, 2.7 +/- .2, and 75.4 +/- 4.6 ng/ml at d 2 to 16.5 +/- 6.5, 1.3 +/- .2, and 53.4 +/- 4.6 ng/ml at d 30, respectively, and remained low through 1 yr of age. Insulin concentration in serum did not change significantly with development. Results indicated that increasing concentrations of specific bGH receptors in the bovine liver may play a key role in regulating postnatal growth in cattle.     

Adaptation of human oscillometric blood pressure monitors for use in dogs

Two digital oscillometric human blood pressure measuring devices were modified and evaluated as blood pressure monitors in 12 healthy anesthetized dogs. Direct arterial pressures were measured via cannulation of the dorsal pedal artery and were correlated with indirect measurements through an inflatable cuff placed over the dorsal pedal artery below the hock joint of the contralateral limb. Direct and indirect measurements were compared for systolic, diastolic, and calculated mean arterial pressures. Blood pressure ranges between 215/145 mm of Hg and 65/30 mm of Hg were obtained, using combinations of halothane, phenylephrine, calcium, and IV administered fluids. Machine A was found to be insufficient for clinical application, on the basis of correlation coefficients between direct and indirect pressures of 0.78, 0.65, and 0.74 for systolic, diastolic, and mean arterial pressures, respectively. Higher correlation coefficients between direct and indirect pressures (0.77, 0.87, and 0.87, respectively) were obtained with machine B. The results of the study reported here suggest machine B may be an effective blood pressure monitoring device in anesthetized dogs.     

Aortic catheterization in cattle via the costoabdominal artery and validation for progesterone and estradiol-17 beta sample collection

The abdominal portion of the aorta was catheterized in 27 cows. Local analgesia was achieved by infiltration of anesthetic agents. A 10-cm skin incision was made caudal and parallel to the 13th rib at the lateral border of the epaxial muscles. The dorsal costoabdominal artery was exposed at its first lateral cutaneous branch by careful dissection through fascial layers. A sterile polyvinyl catheter (1.52 mm OD) was inserted into the artery and was advanced 35 to 40 cm to the abdominal portion of the aorta. Catheter patency was maintained for up to 5 weeks. Concentrations of plasma progesterone and estradiol-17 beta in samples obtained from the abdominal portion of the aorta were similar to simultaneously obtained concentrations in samples from the jugular vein before and after parturition.     

Periparturient endocrine changes of conceptus and maternal units in Holstein heifers bearing genetically different conceptuses

Holstein heifers (n = 21) were balanced across sires and assigned to three service-sire-breed groups in which heifers were inseminated artificially to either purebred Angus (n = 7), Holstein (n = 7) or Brahman (n = 7) bulls. Semen from four bulls was used for each service sire-breed group. Blood samples were collected from a jugular vein thrice weekly from d 160 to 265 of pregnancy, daily thereafter until 15 d postpartum, and then thrice weekly until d 60 postpartum. Concentrations of progesterone, estrone, estradiol, and estrone sulfate from 23 d prepartum to parturition, and of 15-keto-13,14-dihydro-prostaglandin F2 alpha (PGFM) from 2 d prepartum to d 15 postpartum were measured by radioimmunoassay. Heifers within the Brahman-service-sire group had longer gestations (P less than .05) than those of Holstein- or Angus-service-sire groups (285.0 vs 278.7), 279.0 d). Calf birth weight was lower (P less than .05) in Angus- than Holstein- and Brahman-service-sire groups (30.6 vs 36.1, 43.4 kg). Daily trends of prepartum maternal progesterone concentrations were approximately 1 ng/ml lower (P less than .01) in Angus- than Holstein- or Brahman-service-sire groups until luteolysis occurred. Heifers bearing crossbred Angus conceptuses had lower daily trends of prepartum estrogens concentrations (P less than .01), whereas heifers of the Holstein- and, even more dramatically, of the Brahman-service-sire groups had a higher magnitude and greater rise of plasma estrogens concentrations between d -10 and -1 prepartum (less than .01). Postpartum mean concentrations (P less than .05) and response curves of PGFM were lower (P less than .01) in the Angus- than in the Holstein- or Brahman-service-sire groups. Calf birth weights were correlated with least-squares means for maternal concentrations of prepartum estrone (r = .57), estradiol (r = .59) and estrone sulfate (r = .64) and postpartum maternal concentrations of PGFM (r = .56). Functional responses of the conceptus (e.g., estrogens) and maternal units (e.g., progesterone and PGFM) were influenced by conceptus genotype during the periparturient period.     

Levels of serum IgE in response to gastrointestinal nematodes in cattle

Serum IgE concentration has been considered a valuable measurement in parasitic infections, yet little has been reported regarding cattle. This study examines the association of IgE levels of nematode-na?ve Holstein steer calves and the level of gastrointestinal parasitism acquired by grazing irrigated pasture for 30 days. Total IgE levels were determined by enzyme-linked immunosorbent assay (ELISA) specific for bovine IgE on serum collected both before and after exposure to parasite-infected pastures. Following necropsy, parasite loads were determined by direct count from the contents of the abomasum, small intestine, cecum and large intestine; species of Ostertagia and Cooperia being the most common helminths found. Significant increases of IgE in the serum of calves with light infestations were seen, whereas calves with moderate to heavy infestations showed only mild IgE increases. With increased parasite burden, the frequency of increased IgE levels was reduced. Additionally, there appeared to be a seasonal correlation relating the level of serum IgE detected to the number of worms counted and to the course of parasite development.