Showing papers by "University of Veterinary Science published in 2009"

Effect of altrenogest-treatment of mares in late gestation on adrenocortical function, blood count and plasma electrolytes in their foals

Abstract: Summary Reasons for performing study: Mares with compromised pregnancies are often treated with altrenogest to prevent abortion. However, there is only limited information about effects on the foal when altrenogest treatment is continued during final maturation of the fetus. Objectives: To determine effects of altrenogest treatment during late gestation in mares on maturity, haematology changes, adrenocortical function and serum electrolytes in their newborn foals. Methods: Six mares were treated with altrenogest (0.088 mg/kg bwt) once daily from Day 280 of pregnancy until foaling and 7 mares served as controls. Results: Foals born to altrenogest-treated mares had a significantly lower neutrophil/lymphocyte ratio on the first day after birth than control foals (P<0.05). Basal plasma cortisol concentrations immediately after birth were higher in foals of altrenogest-treated mares than in control foals (P<0.05). Cortisol release in response to exogenous adrenocorticotropic hormone (ACTH) - except for higher values 15 min after ACTH injection in foals of altrenogest-treated mares on Day 1 - revealed no differences in adrenocortical function between the groups of foals. Plasma potassium concentration in foals from altrenogest-treated mares compared to control foals was significantly lower immediately after birth (P<0.05) and plasma ionised calcium concentration was significantly lower 3 h after birth (P = 0.01). Conclusions and potential relevance: Altrenogest treatment of pregnant mares prolonged labour had no major effects on adrenocortical function in foals. A reduced neutrophil/ lymphocyte ratio in these foals may suggest either immunomodulatory effects of altrenogest or dysmaturity of the foals.

Cytokines, Growth Factors and Prostaglandin Synthesis in the Uterus of Pregnant and Non-pregnant Bitches: The Features of Placental Sites

Abstract: Uterine tissue from pregnant bitches was investigated by qualitative RT-PCR for the gene expression of local factors potentially important for the implantation of canine embryos. For this purpose, 10 bitches identified as being at the time of implantation or early placentation by means of ultrasonography before ovariohysterectomy (days 20-35, n = 10) provided tissues for comparison to tissue collected in a previous study and identified as early pregnant (n = 10) or non-pregnant (n = 4) by embryo flushing after ovariohysterectomy (days 10-12 after mating; Schafer-Somi et al. 2008). Uterine tissue was excised from the middle of the left horn from early pregnant and non-pregnant animals, including from interplacental and placentation sites. The following genes were investigated: CD-4, -8; cyclooxygenase (COX)-1, -2; granulocyte macrophage-colony stimulating factor (GM-CSF); hepatocyte growth factor (HGF); insulin-like growth factor (IGF)-1, -2; transforming growth factor (TGF) and tumour necrosis factor (TNF)-alpha; interferon (IFN)-gamma; interleukin (IL)-1beta, -2, -4, -6, -8, -10, -12; leukaemia inhibitory factor (LIF) and leptin. Gene expression for CD-8, COX-1, TGF-beta, HGF, IGF-1, IL-2, -4,-10, IFN-gamma and LIF were detected in the pre-implantation uterus, and all except IL-2 and -10 were still detectable during the implantation and placentation stage. During implantation, mRNA for IGF-2 and GM-CSF were additionally detected. The dioestrous uterus differed from the pregnant uterus because of the absence of CD-8, IL-4 and IFN-gamma and the expression of CD-4, TNF-alpha and IL-6. The results suggest that IL-4, IFN-gamma, CD-8, GM-CSF and IGF-2 are regulated in a pregnancy-specific manner and that GM-CSF and IGF-2 probably have growth supporting and immune modulating functions during implantation of the canine embryo.

Expression of MHC‐I and ‐II in Uterine Tissue from Early Pregnant Bitches

Abstract: The aim of the study was to investigate the expression of major histocompatibility complex (MHC)-I and -II in uterine tissues from pregnant and non-pregnant bitches, taken at different time periods after mating. The pregnant bitches were ovariohysterectomized during the pre-implantation (group 1, n = 4), implantation (group 2, n = 7) and placentation stage (group 3, n = 7). Non-pregnant animals in diestrus served as controls (group 4, n = 7). The expression of MHC- I and -II in salpinx, apex, middle horn, corpus uteri and at implantation sites was investigated by immunohistochemistry as well as qualitative and quantitative RT-PCR; MHC-I mRNA was detected in all tissues and with quantitative RT-PCR, and no significant changes were detected until placentation. Immunohistologically, at the apex and corpus site, the average number of MHC-II positive cells increased from the pre-implantation to the post-implantation stage (apex: 1.54 +/- 1.21 to 3.82 +/- 2.93; corpus: 1.62 +/- 1.9 to 5.04 +/- 4.95; p 0.05). Immunohistologically, in the salpinx, significantly greater numbers of MHC-II positive cells were found in the tissues of pregnant animals than in the control group (p < 0.05). It is proposed that the increase in MHC-II is pregnancy-related, even though the impact on maintenance of canine pregnancy is still unclear.

Effect of different concentrations of ascorbic acid on motility, membrane integrity and chromatin status of frozen-thawed canine spermatozoa within six hours of storage at 37°C.

Abstract: The aim of this study was to examine comprehensively the effect of ascorbic acid (Asc) on frozen-thawed canine semen. Pooled ejaculates (n = 10) were assessed for quality with a computer-assisted sperm analyser (CASA). After centrifugation, the semen was divided into four aliquots (A-D) of which three were diluted with Uppsala 1 extender (v/v) containing different concentrations of Asc (B: 6.8 mmol/l; C: 13 mmol/l; D: 27 mmol/l). One group without Asc served as control (A). Subsequently, dilution samples were treated and cryopreserved as described previously (Theriogenology 66, 2006, 173). After thawing, samples were stored at 37 degrees C for 1, 3 and 6 h, then examined for quality (CASA, flow cytometry, sperm chromatin structure assay; SCSA). Staining for flow cytometry was performed with FITC-PNA and propidium iodide (Reproduction 128, 2004, 829). The SCSA was performed with both Tris-NACL-EDT buffer (TNE)- and Tris-fructose-citrate buffer (TFC). In the Asc-supplemented groups, percentages of progressively motile sperm (P) were significantly lower than in the control group (A 1 h: 56.6 +/- 9.8, 6 h: 18 +/- 4.5; B 1 h: 51.2 +/- 11.8, 6 h: 13.6 +/- 5; C 1 h: 41 +/- 16.2, 6 h: 11.2 +/- 6.1; D 1 h: 37 +/- 15.2, 6 h: 8.6 +/- 5; p 0.05). Furthermore, there were no significant differences between TNE- and TFC-samples, for alphaT, for SD of alphaT or for comp alphaT [comp alphaT: TCF-A: 2.5 +/- 1.7%, TNE-A: 3.4 +/- 3.2%; TCF-D: 2.5 +/- 2%, TNE-D: 3.3 +/- 4.3%; p > 0.05]. However, samples diluted with both extenders correlated concerning alphaT, but not comp alphaT. We therefore recommend using TNE-buffer for SCSA with cryopreserved canine semen. In addition, the average alphaT values did not differ significantly between the controls and all other groups (TNE-A: 380.2 +/- 89.1; TNE-D: 338.1 +/- 137.4; p > 0.05). It can be concluded that addition of Asc to cryoextender does not increase quality of frozen-thawed canine semen.

Protein quality of mixtures of feed ingredients

Abstract: Zusammenfassung Proteinqualitat von Futtermittelmischungen II. Weizen und Schlachtabfallmehle Die Proteinqualitat von Mischungen aus Weizen und Mehlen von Schweine- und Geflugelschlachtabfallen wurde mit Hilfe folgender Kriterien gepruft: NPU-Wert, Korpergewichtsentwicklung und Futterverwertung bei der Ratte, Aminosaurezusammensetzung, limitierende essentielle Aminosaure, Summe der essentiellen Aminosauren, Chemical score, PV-Wert und EAA-Index. Es wurden 0; 1; 2; 3; 4; 6; 8; 10 und 20% des Weizens durch tierisches Proteinfutter ersetzt. Bei den in vivo-Untersuchungen wurde in jeder Mischung ein Erganzungseffekt nachgewiesen. Der Chemical score und EAA-Index zeigten Erganzungswirkungen nur bei Weizen und Schlachtmehlen vom Schwein. Aus der Summe der essentiellen Aminosauren liesen sich keine Voraussagen fur den NPU-Wert bei der Ratte ableiten. Die Korrelationskoeffizienten zwischen in vivo- und in vitro-Methoden hangen von Art und Mischungsverhaltnis der beiden Komponenten ab.

68 vitrification of cleavage-stage mouse embryos by the vitroloop (cryoloop) procedure

Abstract: By decreasing the volume of the cryoprotective solution, we were able to dramatically increase the freezing speed and decrease the toxicity and osmotic side effects of the cryoprotectants (CP). Several carriers have been developed successfully (Vajta G et al. 1998 Mol. Reprod. Dev. 51, 53–58; Liebermann J et al. 2002 Reprod. Biomed. Online 4, 146–150; Park SP et al. 1999 Hum. Reprod. 15, 1787–1790; Chung HM et al. 2000 Fertil. Steril. 73, 545–551; Kuwayama M and Kato O 2000 Fertil. Steril. 74(Suppl. 3), S49 O-127; Matsumoto H et al. 2001 Cryobiology 42, 139–144; Lane M et al. 1999 Fertil. Steril. 72, 1073–1078; Dinnyes A et al. 2000 Biol. Reprod. 63, 513–518). The objective of our study was to vitrify Day 3 cleavage stage mouse embryos with the Vitroloop™ (Vitrolife, Kungsbacka, Sweden) cryopreservation technology. Vitrification was carried out in RapidVit™ Cleave (Vitrolife) solutions (holding, equilibration, and vitrification medium). Embryos were exposed to a 2-step loading of CP, ethylene glycol (EG), and propylene glycol (PG), before being placed in a small loop attached to the lid of a cryo-vial and rapidly submerged into liquid nitrogen (LN). First, the embryos were transferred from the G-MOPS holding medium to the equilibration medium containing 8% EG for 2 min. Then, embryos were transferred into a 20-μL drop of vitrification medium containing 16% EG, 16% PG, 10 mg mL–1 Ficoll 400, and 0.65 m sucrose for 30 s. After that, the embryos (maximum of 2 at a time) were transferred onto the loop, which was quickly sealed in a cryo-vial of LN and stored. After storage in LN, embryos were warmed by a 3-step dilution of the CP with sucrose (RapidWarmCleave™, Vitrolife) carried out at 37°C. First, the loop with the embryos was quickly immersed in 37°C warming medium 1 (0.65 m sucrose) for 20 s. Then, the embryos were transferred into warming medium 2 (0.25 m sucrose) for 1 min, then into warming medium 3 (0.125 m sucrose) for 2 min, and finally into warming medium 4 (G-MOPS medium) for 5 min. Following warming, embryos were cultured in G1 medium (Vitrolife) at 37°C with 6.5% CO2 and maximum humidity in air. Embryo viability was assessed by 48 h in vitro culture; the survival of embryos was based on morphological appearance in vitro after thawing and continued development to expanded blastocysts upon subsequent culture. The control embryos were treated likewise except that they were not vitrified. A total of 229 cleavage-stage embryos were vitrified and warmed; out of these, 11 were lost (11/229; 4.8%). Of the remaining 218 embryo, 202 survived vitrification (202/218; 92.7%) and 180 developed further to expanded blastocysts during in vitro culture (180/202; 82.6%). In the control group, 91.4% of the embryos developed to expanded blastocysts (75/82) indicating that the solutions used were not toxic. Our data show that a high percentage of cleavage-stage mouse embryos survived vitrification in the mixture of EG and PG combined with the use of cryoloop and developed normally in vitro after thawing. To our knowledge, this is the first report of the successful use of the Vitroloop™ vitrification procedure with cleavage-stage mouse embryos. The authors thank Vitrolife Ltd. (Kungsbacka, Sweden) and FertiCad Ltd. (Budapest, Hungary) for providing the solutions. The 3-month fellowship for Phillip Klambauer was provided by CEEPUS.