bluuebunny/crossref_metadata_2025_split huggingface dataset:
Stimulation of Mating‐Induced Uterine Contractions in the Bitch and Their Modification and Enhancement of Fertility by Prostatic Fluid
Authors: England GCW , Moxon R , Freeman SL 
Published: December 2012

ContentsLittle is known about the response of the bitch's reproductive tract to mating or of the role of male accessory gland secretions in the female. In this clinical study, the component stimuli causing mating‐induced uterine contractions were investigated in 64 bitches. Basal uterine contractions were present during oestrus and a significant increase in the frequency of contractions was observed during natural mating. Neither teasing with a male nor stimulation of the vagina or cervix by vaginal or transcervical insemination (TCI) caused an increase in the frequency of uterine contractions. Increased contractions were however present after both vaginal and transcervical insemination when the vestibule was distended, and dorsal wall of the vaginal was manually stimulated. Interestingly, this increase in uterine contractions was partially ameliorated when prostatic fluid was used as a flushing component following transcervical insemination. Two further studies performed with 72 bitches of which 18 were each inseminated transcervically with fresh or frozen semen flushed into the uterus with either saline or prostatic fluid demonstrated that prostatic fluid significantly increased the pregnancy rate and litter size of both groups. There are important mechanisms regulating the transport and elimination of sperm from the bitch's reproductive tract. Whilst physical aspects of coitus are undoubtedly involved in initiating uterine contractions, prostatic fluid appears to have an important role in modulating uterine contractions and fertility.
Customizing Semen Preservation Protocols for Individual Dogs and Individual Species: Sperm Preservation beyond the State of the Art
Authors: Farstad W 
Published: December 2012

ContentsSperm quality can be variable in morphometric and physiological attributes between males of different species, between males within species subtypes reared under different environmental conditions, between ejaculates of the same male or even between sperm populations within an ejaculate. Clinical semen evaluation is based on evaluation of whole ejaculates, which is not a chemically or physiologically well‐defined entity, rather a collection of heterogeneous subpopulations giving different measurements and possessing different fertilizing potential. Identification of subpopulations with different motility patterns is important as well as characterizing the subtle structural changes underlying the motility differences observed. The ability to identify populations of sperm responding rapidly or failing to progress through the capacitation process may have clinical applications. Studies of lipid‐phase fluidity of sperm membranes, mathematical modelling of membrane ion transport, role of modifying components and detergent‐resistant microdomains are of particular interest. When customizing extenders to ejaculates from cryosensitive males or species, a thorough knowledge of species sperm membrane physiology and an assessment of the individual ejaculate's sperm populations are necessary. Structural differences have been found in sperm membranes between fox species with different cryosurvival potential of their spermatozoa. Supplementation of lipids and detergents in cryoextenders may influence membrane fluidity of the surviving spermatozoa in a species‐dependent manner and influence capacitation. Immobilization of sperm prior to cryopreservation with subsequent slow release of sperm in the female genital tract may be a way to prolong the fertile life of sperm. In canids with a long oocyte maturation time, delayed capacitation may be beneficial.
Gene Expression in Pre‐Term, Pre‐Labour and Parturient Canine Placenta
Authors: Fellows EJ , Hazzard TM , Kutzler MA 
Published: December 2012

ContentsMatrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), vascular endothelial growth factor (VEGF)‐A, VEGF‐A receptor (Flt‐1) and KiSS‐1 receptor (KiSS‐1R) all play a role in trophoblast invasion in a number of mammalian species. However, mRNA expression of factors regulating trophoblast invasion has not been studied in dogs. Abnormal expression of these factors at the end of canine gestation may contribute to placental retention and/or subinvolution of placental sites. Therefore, we sought to determine the relative mRNA expression of these factors in canine chorioallantois tissue at 61 ± 1 day past the LH surge (pre‐term; n = 4), following elective c‐section at 64 ± 1 day past the LH surge prior to first‐stage labour (pre‐labour; n = 4) and following natural delivery (parturient; n = 3). Total RNA was isolated, real‐time RT‐PCR was performed, and relative expression was calculated using the relative quantitation () method. MMP‐9 mRNA expression was significantly higher in pre‐term samples compared to pre‐labour and parturient samples. The results showed no significant difference between MMP‐2, TIMP‐2, VEGF‐A and Flt‐1 mRNA expression among the three groups. KiSS‐1R mRNA was not expressed in any tissues studied. Gene expression of MMP‐9 may be related to the onset of labour, whereas MMP‐2, VEGF‐A, Flt‐1, TIMP‐2 and KiSS‐1R mRNA do not appear to play a role at the end of gestation in the dog.
Cat and Dog Primordial Follicles Enclosed in Ovarian Cortex Sustain Viability after In vitro Culture on Agarose Gel in a Protein‐Free Medium
Authors: Fujihara M , Comizzoli P , Wildt DE , Songsasen N 
Published: December 2012

ContentsOur objective was to examine the influences of differing media, protein supplementation and the microenvironment on cat vs dog primordial follicle viability in vitro. Ovarian cortical slices were cultured for 3, 9 or 15 days in α‐minimum essential medium (α‐MEM) or MEM supplemented with 10% fetal bovine serum (FBS), 10% knock‐out serum replacement (KSR) or 0.1% polyvinyl alcohol (protein free). In a separate study, cat and dog ovarian tissues were cultured in protein‐free α‐MEM and MEM, respectively, in cell culture inserts, on 1.5% agarose gel or in 24‐well cell culture plates (control). Follicle viability was assessed in both studies using calcein AM/ethidium homodimer and histological evaluation with haematoxylin/eosin staining. No cat follicle sustained viability beyond 9 days of in vitro culture in α‐MEM compared to 37.5% of those incubated for 15 days in MEM in protein‐free condition (p < 0.05). In contrast, α‐MEM was superior (p < 0.05) to MEM in maintaining dog follicle viability (32.7% vs 8.1%) in protein‐free condition at 15 days. Serum was detrimental (p < 0.05) to follicle survival in both species. Knock‐out serum replacement supplementation and a protein‐free condition supported cat follicle viability, whereas the latter was superior (p < 0.05) to the former for sustaining dog follicle survival. Likewise, dog follicle viability was enhanced (p < 0.05) by the agarose gel compared to the cell culture insert and control groups after 3 and 9 days of culture. For the cat, the agarose gel better (p < 0.05) supported follicle viability compared to the control, but was equivalent to the cell culture insert. Therefore, sustaining primordial follicle survival from intracortical ovarian slices requires a different in vitro microenvironment for the cat vs the dog. A key factor to enhancing survival of these early stage follicles in culture appears to be the use of agarose gel, which enhances follicle viability, perhaps by promoting gas exchange.
Efficacy of Cloprostenol or Aglepristone at 21–22 and 35–38 Days of Gestation for Pregnancy Termination in Queens
Authors: García Mitacek MC , Stornelli MC , Praderio R , Stornelli MA , de la Sota RL 
Published: December 2012

ContentsThe objective of this study was to evaluate the efficacy of cloprostenol (CLO) or aglepristone (ALI) for pregnancy termination in queens at 21–22 and 35–38 days of gestation. Two experiments (EXP) were carried out to accomplish this aim. Thirty‐seven 12‐ to 14‐month‐old mixed breed queens were used in a randomized design. At oestrus, queens were housed for mating with a tom, and pregnancy was confirmed by transabdominal ultrasonographic examination (US). On days 21–22 of pregnancy (EXP1) or 35–38 of pregnancy (EXP2), queens were divided into three groups (G). Queens in G1 received ALI (10 mg/kg, sc; EXP1, n = 6; EXP2, n = 6) on two consecutive days. Queens in G2 received CLO (5 μg/kg, sc; EXP1, n = 6; EXP2 = 7) on three consecutive days. Queens in G3 received 1 ml of saline solution (PLA, sc; EXP1, n = 6; EXP2 = 6). After treatment, females were monitored daily by US during for 10 days and weekly until the end of gestation. In EXP1, pregnancy was terminated in (6/6, 100%), (0/6, 0%) and (0/6, 0%), for the ALI, CLO and PLA groups, respectively (p < 0.001). In EXP2, pregnancy was terminated in (6/6, 100%), (1/7, 14%) and (0/6, 0%) for the ALI, CLO and PLA groups, respectively (p < 0.001). In both EXP, after CLO administration, animals vomited and were depressed for 30 min; but no side effects were observed in the animals in the ALI group. In conclusion, the results from this study indicate that three injections of CLO are not effective, but two injections of ALI are effective to induce abortion in queens at 21–22 or 35–38 days of pregnancy.
Unilateral Orchidectomy in Mature Cats is Not Followed by Compensatory Hypertrophy
Authors: Romero GG , Barbeito C , Fernandez P , Gimeno E , Gobello C 
Published: December 2012

ContentsThe aim of this study was to describe the effect of unilateral orchidectomy on testicular characteristics of mature domestic cats. Five, 1‐ to 2‐year‐old, cross‐bred male cats were unilaterally orchidectomized on day 0 (right) and day 60 (left). The testes were processed for histological and immunohistochemical evaluation and groups compared by Student's t‐test. No significant differences (p > 0.05) were found between testes groups for any of the gross (volume and gonadosomatic index) and microscopic (tubular diameters, axis, area, length and perimeter, germinal epithelium height, spermatogonias, spermatocytes, spermatids, spermatozoa, Sertoli and Leydig cells, intertubular compartment, lumen, cellular debris, tubular‐ intertubular proportion and basement membrane) parameters assessed. According to these biometric and morphometric results, mature cats do not develop compensatory hypertrophy within 60 days of unilateral orchidectomy.
Effects of GnRH Antagonists vs Agonists in Domestic Carnivores, a Review
Authors: Gobello C 
Published: December 2012

ContentsGonadotrophin‐releasing hormone (GnRH) stimulates the pituitary secretion of both luteinizing and follicle‐stimulating hormones, and thus controls the hormonal and reproductive functions of the gonads. GnRH analogs, which include agonists and antagonists, have been produced by amino acid substitutions within the native GnRH molecule resulting in greater potency and a longer duration of effectiveness. While the initial antagonists produced significant side effects, more recent potent, long‐acting, water‐soluble, low histamine‐release third‐generation compounds such as cetrorelix, abarelix, azaline B and acyline have appeared. Differently to GnRH agonists, antagonists competitively block and inhibit GnRH‐induced GnRH receptor gene expression leading to an immediate, dose‐dependent, pituitary suppression without an initial stimulation of the gonadal axis. The aims of this review are to compare the effects of GnRH agonists vs antagonists and to describe the existing literature concerning new antagonists in domestic carnivores. In male dogs, a single subcutaneous dose of acyline safely and reversibly decreased serum gonadotrophins and testosterone concentrations for 9 days and prevented physiological response of gonadal the axis to agonistic challenge for 14 days. The same protocol reversibly impaired spermiogenesis, spermatocytogenesis and semen quality in both cats and dogs. In females, third‐generation GnRH antagonists prevented ovulation and interrupted pregnancy in canids but not in felids. During anestrus, a single acyline injection exhibited limited prevention of the ‘flare‐up’ effect in GnRH agonist‐implanted bitches. Although GnRH antagonists appear to have a promising future in domestic carnivores reproduction, the information is still scarce and further work is needed before they can be widely recommended.
Embryo Retrieval after Hormonal Treatment to Control Ovarian Function and Non‐surgical Artificial Insemination in African Lions (Panthera leo)
Authors: Goeritz F , Painer J , Jewgenow K , Hermes R , Rasmussen K , Dehnhard M , Hildebrandt TB 
Published: December 2012

ContentsAssisted reproduction technologies are essential for propagating endangered wild felids. Artificial insemination (AI) has been reported in several wild feline species, but pregnancy rates are low, partially owing to failures of current hormonal stimulation protocols. Therefore, this study describes the application of reliable methods to monitor ovarian activity and the development of an effective hormonal protocol to induce oestrus and ovulation in African lions. Application of porcine FSH and porcine LH was shown to be effective for inducing follicular growth and ovulation, and this regimen appeared to be superior to protocols described earlier in terms of ovulation and fertilization rates. Furthermore, non‐surgical AI was performed successfully in lions, and uterine‐stage embryos were collected and cryopreserved. African lions may serve as a valuable model to develop assisted reproduction for propagation of relic zoo populations in the critically endangered Asian lion or Barbary lion.
Scriptaid and 5‐aza‐2′deoxycytidine Enhanced Expression of Pluripotent Genes and in vitro Developmental Competence in Interspecies Black‐Footed Cat Cloned Embryos
Authors: Gómez MC , Biancardi MN , Jenkins JA , Dumas C , Galiguis J , Wang G , Earle Pope C 
Published: December 2012

ContentsSomatic cell nuclear transfer offers the possibility of preserving endangered species including the black‐footed cat, which is threatened with extinction. The effectiveness and efficiency of somatic cell nuclear transfer (SCNT) depends on a variety of factors, but ‘inappropriate epigenetic reprogramming of the transplanted nucleus is the primary cause of the developmental failure of cloned embryos. Abnormal epigenetic events such as DNA methylation and histone modifications during SCNT perturb the expression of imprinted and pluripotent‐related genes that, consequently, may result in foetal and neonatal abnormalities. We have demonstrated that pregnancies can be established after transfer of black‐footed cat cloned embryos into domestic cat recipients, but none of the implanted embryos developed to term and the foetal failure has been associated to aberrant reprogramming in cloned embryos. There is growing evidence that modifying the epigenetic pattern of the chromatin template of both donor cells and reconstructed embryos with a combination of inhibitors of histone deacetylases and DNA methyltransferases results in enhanced gene reactivation and improved in vitro and in vivo developmental competence. Epigenetic modifications of the chromatin template of black‐footed cat donor cells and reconstructed embryos with epigenetic‐modifying compounds enhanced in vitro development, and regulated the expression of pluripotent genes, but these epigenetic modifications did not improve in vivo developmental competence.
Characterization of Mitochondrial and Actin Patterns in Cat Oocytes and Blastocysts
Authors: González R , Gómez MC , Pope CE , Brandt YCB 
Published: December 2012

ContentsActin microfilaments and mitochondria distribution are considered useful markers of cytoplasmic maturation, but no information is available regarding their distribution in cat oocytes and embryos. Thus, the purpose of this study was to (i) assess cytoplasmic characteristics of the oocyte by mitochondria and actin staining in immature and in vitro/in vivo matured cat oocytes and (ii) characterize mitochondria and actin distribution in in vitro produced blastocysts by confocal laser scanning microscopy. Additionally, in vivo matured oocytes were collected to assess mitochondria and actin. Transzonal cumulus cell projections were more abundant in immature oocytes than in matured oocytes. A relocation of mitochondria throughout meiosis was not clearly observed. However, most in vitro produced blastocysts were of good quality, according to their actin cytoskeleton integrity and mitochondria distribution. The functional significance of mitochondria distribution in cat oocytes in relation to their developmental competence requires further research. This study represents the original description of actin and mitochondrial patterns in cat oocytes and embryos.
Serum Progesterone in Pregnant Bitches Supplemented with Progestin Because of Expected or Suspected Luteal Insufficiency
Authors: Günzel‐Apel A , Urhausen C , Wolf K , Einspanier A , Oei C , Piechotta M 
Published: December 2012

ContentsProgesterone profiles of individual bitches may vary considerably both between and within individuals during pregnancy and non‐pregnancy. Suspected luteal deficiency is commonly purported but is difficult to evaluate in clinical cases when progesterone is supplemented because this masks the underlying hormone changes. Therefore, in this study, suspected cases of luteal deficiency (six pregnancies from five bitches) were supplemented with oral medroxyprogesterone acetate (MPA), allowing measurement of endogenous progesterone using conventional assay. MPA (0.1 mg/kg) treatment commenced between days 30 and 36 after estimated ovulation and was continued for 18–28 days. Endogenous progesterone was measured throughout treatment, and blood was additionally analysed for prolactin (PRL) and relaxin (RLN) as well as MPA. The latter revealed delayed MPA clearance in two bitches, in which Caesarean operation had to be performed because of a low foetal heart rate. In two cases with confirmed basal concentrations of both P4 and MPA at term, spontaneous parturition occurred. Low endogenous progesterone during pregnancy was not apparent in three bitches that had previously had a short inter‐oestrous interval of which two had previously had confirmed short luteal phase. However, in the remaining two cases, there had been previous pregnancy failure, but in only one of these, a premature decrease in endogenous progesterone to <2 ng/ml was detected. The latter had also low concentrations of PRL and RLN. The results of this preliminary clinical study suggest that abnormal progesterone profiles in pregnancy may be uncommon in bitches even when there has been previously documented short inter‐oestrous interval. However, luteal deficiency may be suspected in bitches with a history of repeated pregnancy failure or abortion. MPA supplementation appears to be efficacious for management of suspected luteal deficiency and verification of the ovarian dysfunction, but care should be taken regarding the timing of MPA withdrawal and prolongation of pregnancy because of delayed elimination of MPA from blood circulation.
Recombinant Feline Oviductin – A Powerful Tool for Functional IVF Studies in the Domestic Cat
Authors: Hachen A , Jewgenow K , Krause E , Braun BC 
Published: December 2012

ContentsOviductins are highly conserved proteins that seem to play an important role during fertilization and embryo development. A universal characteristic of oviductin is its association with the zona pellucida of ovulated oocytes, and several functional studies indicate an influence of this glycoprotein on reproductive events in mammals. The objective of this study was to produce recombinant feline oviductin in prokaryotic as well as eukaryotic cells as a prerequisite for analysing its influence on IVF in the domestic cat. Oviductin sequence was cloned into pET21b vector and transformed to E. coli to produce a recombinant His‐tagged, non‐glycosylated protein. Oviductin was isolated from E. coli lysate using anion exchange chromatography followed by immobilized metal ion affinity chromatography (IMAC). Western blot analysis of affinity purified fractions resulted in one clear band corresponding to the expected size of ~67 kDa. The corresponding band of a coomassie‐stained gel was analysed by mass spectrometry. Oviductin was identified with over 50 peptides covering 72% of the whole protein sequence. To obtain a glycosylated form of oviductin, eukaryotic cells were stable transfected with pSecTag/HygroA vector containing the oviductin gene sequence. The recombinant His‐tagged protein was harvested from a serum‐ and protein‐free cell culture medium. Mass spectrometry analyses of protein bands obtained after separation of the medium by SDS‐PAGE detected oviductin peptides in protein bands of ~70, ~85 and ~170 kDa. With prokaryotic as well as eukaryotic produced recombinant feline oviductin, we are now able to use the protein for further functional IVF studies in the domestic cat.
Clinical and Molecular Characteristics of Pyometra in Female Dogs
Authors: Hagman R 
Published: December 2012

ContentsPyometra is a common disease of female dogs. In Sweden, where approximately 90% of the dog population is intact (not neutered), nearly 25% of all female dogs are diagnosed with the disease before 10 years of age. In certain high‐risk breeds, this risk of developing pyometra exceeds 50%. Various clinical signs associated with the genital tract as well as with systemic disease are present in dogs with pyometra. A frequent and serious consequence of the uterine infection is endotoxaemia and progression into the systemic inflammatory response syndrome (SIRS), or sepsis, and the disease is then regarded as a medical emergency. Acute phase proteins and inflammatory markers associated with SIRS and with the outcome as measured by length of hospitalization have been identified in blood samples. Recently, the inflammatory response in infected uterine tissue during pyometra has been more closely explored. The expression of many genes associated with chemokines, cytokines, inflammatory cell extravasation, anti‐bacterial action, the complement system and innate immune responses and also a large panel of proteases are upregulated in the uterine tissue in pyometra. Products of certain upregulated genes may be detected systemically and used for diagnostic or prognostic purposes provided that tests are developed in the future. More knowledge of the complex local and systemic inflammatory response in pyometra may allow identification of novel disease biomarkers or future targets for treatment. In this article, clinical as well as molecular characteristics of the disease are reviewed.