bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison Between Vestibular and Subcutaneous Insertion of Deslorelin Implants for Oestrus Induction in Bitches
Authors: Kutzler M , Lamb SV , Volkmann D 
Published: July 2009

ContentsInvestigations using sustained‐release deslorelin implants at various insertion sites have shown that this method consistently induces oestrus in anoestus bitches. However, fertility comparisons between implant insertion sites have not been performed. Anestrous beagle bitches received one 2.1 mg deslorelin implant beneath the vestibular mucosa (VM group; n = 6) or in the subcutaneous tissue between the shoulder blades (SubQ group; n = 8). Vestibular implants were removed when serum progesterone concentrations first exceeded 1.5 ng/ml. Vaginal cytologies and blood samples were collected daily and bitches were inseminated during oestrus. Serum progesterone and deslorelin concentrations were measured and pregnancy status was determined using ultrasonography. There were no differences between groups in the intervals between implant administration and the onset of proestrus, the time of the luteinizing hormone surge and the onset of cytologic diestrus. There were also no differences in the number of corpora lutea or foetuses. However, conception rate was significantly lower in the SubQ group. The pregnancy rate did not differ significantly between the VM and SubQ groups [4 out of 6 (66.7%) and 3 out of 8 (37.5%), respectively]. One bitch (16.7%) in the VM group and three bitches (37.5%) in the SubQ group suffered distinct, premature declines in serum progesterone concentrations starting 1–4 weeks after cytologic diestrus. Serum progesterone concentrations did not recover (premature luteal failure), resulting in abortion. Bitches with premature luteal failure in the SubQ group still had serum deslorelin concentrations >100 pg/ml 20 days after implant insertion, suggesting a possible association between prolonged deslorelin release and luteal failure.
Prostaglandin Synthesis Genes are Differentially Transcripted in Normal and Pyometra Endometria of Bitches
Authors: Silva E , Leitão S , Ferreira‐Dias G , Lopes da Costa L , Mateus L 
Published: July 2009

ContentsPro‐inflammatory stimuli, such as endotoxins released by Gram‐negative bacteria, are potent stimulators of prostaglandin (PG) synthesis. The aim of this study was to evaluate the gene transcription pattern of PG synthesis enzymes in normal (anestrous, n = 6 and diestrous, n = 8) and pyometra (n = 7) endometria of bitches. Uteri were collected during routine ovariohysterectomy, processed for histopathological evaluation and uterine contents cultured. Gene transcription of COX‐1, COX‐2, mPGES‐1 and PGF‐synthase (PGFS) were evaluated by relative real‐time PCR and normalized with the ribosomal protein L27 (RPL27) housekeeping gene. Normal uteri had no histological abnormalities and were negative for bacteriology. All pyometra uteri were hyperplasic and Escherichia coli was the only isolated bacterium. Except for COX‐1, gene transcription was significantly higher in pyometra than in normal endometria. No significant differences in gene transcription were observed between normal diestrous and anestrous endometria. COX‐2 gene transcription was 19 and 69 times higher in pyometra than in diestrous and anestrous endometria (p < 0.001), while PGFS gene transcription had a 3‐ and 600‐fold increase in pyometra endometria compared to normal diestrous and anestrous endometria (p < 0.001). Gene transcription of mPGES‐1 was 9 times higher in pyometra than in normal uteri (p < 0.01). Based on these results, we suggest that pyometra‐associated E. coli endotoxin release stimulates the up‐regulation of COX‐2 PGFS and mPGES‐1 gene transcription in the endometrium.
Expression Pattern of Markers in the Canine Ovarian Cycle
Authors: Thuróczy J , Oppe N , Scheuermann E , Perge E , Tibold A , Balogh L 
Published: July 2009

ContentsAlthough hormonal changes during different phases of the oestrous cycle of bitches are well‐described, knowledge about the luteal phase and anoestrus is incomplete. Furthermore, which paracrine and autocrine critical factors that differentiate between follicles destined for atresia and those that continue to develop are unknown. In this study, ovarian tissue was collected from 39 healthy bitches that were subject to ovariectomy or ovariohysterectomy for surgical neutering or medical purposes such as unwanted pregnancy. Bitches were allocated to different groups depending on the stage of the oestrous cycle. Serum progesterone, LH, FSH and 17β‐estradiol (E2) ‐levels were determined and immunhistochemistry was performed for a variety of receptor antigens; Ki‐67, vimentin, pan cytokeratin antibody, p53 and oestrogen receptor (ER) α antigens. Marked differences were found in progesterone concentration between pregnant and non‐pregnant animals. Oestrogen concentration was significantly lower in pro‐oestrus and ovulation than during the luteal phase. Although progesterone could be detected in cytoplasm of ovarian cells at each stage, its presence was restricted to follicular cells during anoestrus. A strong presence of AE1/AE3, vimentin and p53 was found in each oestrous stage, in contrast with Ki67. The localization of ERα appeared to vary during the oestrous cycle, a phenomenon that suggests a switch between target cells of oestrogen; while as a proliferation marker, the mild reaction of p53 during parturition suggests an apoptotic process at this stage of the cycle.
Progesterone, Oestradiol, FSH and LH Concentrations in Serum of Progesterone‐Treated Pregnant Bitches with Suspected Luteal Insufficiency
Authors: Tibold A , Thuróczy J 
Published: July 2009

ContentsIn the bitch, the corpus luteum is the only source of circulating progesterone throughout pregnancy. Inadequate luteal function may be a cause of abortion or foetal resorption observed after early pregnancy diagnosis. In our study of factors involved in canine luteal inadequacy, 35 pregnant bitches from different breeds were allocated to groups of healthy control (n = 15) vs hypoluteoid (n = 20) pregnant bitches, based on presence or absence of clinical signs of impending abortion and on progesterone concentrations below 10 ng/ml at the 4th–5th week of pregnancy. Hypoluteoid bitches were treated daily with 10‐mg natural progesterone in injectable form (Luteosan® inj.; Alvetra and Werfft AG, Vienna, Austria) until day 60. Serum progesterone, 17β‐oestradiol, FSH and LH concentrations were measured in samples obtained weekly using ELISA previously validated for dog serum. The exogenous progesterone supplementation was presumed to be sufficient to prevent foetal loss in 15 of the 20 treated bitches. The mean serum progesterone concentration in control pregnant bitches did not decrease below 10 ng/ml until the 8th week of pregnancy. Progesterone concentrations in progesterone‐treated hypoluteoid bitches at week 4 were lower than in controls (p < 0.05), but although numerically higher did not differ significantly from those of controls during the period of treatment. Serum 17β‐oestradiol concentrations of healthy bitches were variable, were at most time higher than those of treated animals and slowly decreased until parturition; those of treated bitches remained unchanged during the study. Serum FSH and LH concentrations did not differ between groups. Additional studies involving untreated pregnancies showing equivalent evidence of hypoluteoidism as well as assay of circulating relaxin and prolactin in treated and untreated bitches are needed to better determine the causes and effects of hypoluteoidism in pregnant bitches.
Comparison of Two Doses of the GnRH Antagonist, Acyline, for Pregnancy Termination in Bitches
Authors: Valiente C , Corrada Y , De La Sota PE , Blanco PG , Arias D , Gobello C 
Published: July 2009

ContentsGonadotropin‐releasing hormone (GnRH) antagonists are particularly useful when a rapid inhibitory effect on the gonadal axis is required. The aim of this study was to test the efficacy and clinical safety of a low and high dose of the third generation GnRH antagonist, acyline, on pregnancy termination in female dogs. The effect of the antagonist on the progesterone (P4) serum concentration was also described. Twenty‐one mid‐pregnant bitches were randomly assigned to a single subcutaneous (SC) dose of a placebo (PLACE; n = 7), a low (ACY‐L; 110 μg/kg; n = 6) or high (ACY‐H; 330 μg/kg; n = 8) dose of acyline. The animals were followed up for 15 days. All ACY treated but no placebo‐treated animals terminated their pregnancy by abortion (p < 0.01). The ACY‐L and ACY‐H groups interrupted their pregnancy 7 ± 1.9 and 6.4 ± 1.3 days after treatment, respectively (p = 0.7). A significant interaction between treatment and day was found (p < 0.01) for P4 serum concentrations when PLACE was compared with both ACY groups. No difference was found for this hormone between both ACY groups (p > 0.05) where P4 diminished throughout the study. The decreasing rate varied among animals and was closely related to the time of abortion when P4 reached basal concentrations. In PLACE animals, gestation progressed normally and P4 did not change throughout the study (p > 0.05). None of the bitches presented side effects. It was concluded that acyline safely terminated mid‐pregnancy by permanently decreasing P4 serum concentrations.
Dystocia in Numbers – Evidence‐Based Parameters for Intervention in the Dog: Causes for Dystocia and Treatment Recommendations*
Authors: Münnich Andrea , Küchenmeister Uwe 
Published: July 2009

ContentsDystocia is a frequent problem in the dog, with regard to breed‐specific factors, litter size, duration of expulsion, various intervals between puppies and various causes of maternal, foetal or combined origin. In the present study, results were obtained from management of 530 dogs of 54 breeds, between 1 and 13 years of age, presented with dystocia over a period of 8 years in an obstetrical clinic with mostly identical diagnostic and therapeutic schedules. Data on age of the dam, parturition number, breed size, causes of dystocia, duration of expulsion time, kind of obstetrical aid, and last but not least, the influence of all therapeutic aspects on survival of the puppies were analysed. Dams of miniature and small breeds (59.4%) had a high incidence for dystocia in this population. Uterine forces (inertia and spasm), malpresentation of the foetus and litter size (single pup pregnancies, hyperfoetation) were the most common causes for dystocia. Duration of expulsion stage had the highest influence on puppy survival (p < 0.001). In the case of medical treatment, medication with oxytocin led to higher puppy losses compared with other medicaments. Older primiparous bitches (>6 years of age) had a significantly higher risk to have special obstetric conditions and stillbirths compared with young primiparous bitches. The scope of this study was to analyse the parameters that were most useful for a rapid diagnosis to maximize puppy survival, to avoid false diagnosis and to choose the best therapeutic strategy. The present data show that in addition to the established criteria for management of dystocia in the dog, new diagnostic approaches are necessary to improve obstetrical care.
Preliminary Study in Immature Canine Oocytes Stained with Brilliant Cresyl Blue and Obtained From Bitches with Low and High Progesterone Serum Profiles
Authors: Rodrigues BA , Rodriguez P , Silva AEF , Cavalcante LF , Feltrin C , Rodrigues JL 
Published: July 2009

ContentsThis study was conducted: (i) to observe the features and levels of blue colour impregnation in morphologically selected immature canine cumulus oocyte complexes (COCs) stained with the brilliant cresyl blue (BCB) dye, as indicators of quality, and integrity of nuclear oocyte chromatin configuration before in vitro maturation (IVM); (ii) to observe the relationship between the influence of serum progesterone (SP) concentrations from ovary donors and BCB staining of immature dog oocytes. The results showed that out of 138 canine COCs, germinal vesicle (GV) stage prevailed in BCB+ oocytes at percentages of 67.4% (60/89), which were statistically higher than those observed in BCB+/− (52.2%; 23/44) and BCB− (20%; 1/5) oocytes (p = 0.023). Oocytes BCB+ were interpreted as those having completed their growth and therefore possessing the capacity to mature and develop in vitro. Ooplasm and cumulus cells (CCs) of canine oocytes were BCB staining independent. Ooplasm blue colour staining reaction varied between grown oocytes, revealing different levels of glucose‐6‐phosphate dehydrogenase activity among and within oocytes. Additionally, SP profile of ovary donors was not a relevant indicator for selection of oocytes screened with the BCB stain. Similar numbers of high quality oocytes were observed to be BCB+, BCB+/− and BCB− between groups of females with SP varying from 0 to 2.5 ng/ml (n = 5), and those with SP varying from 2.6 to 16.7 ng/ml (n = 4) (p = 0.680). It may be inferred that bitches with low and high SP profiles have grown oocytes in their ovaries, as determined by the BCB absorbance in their ooplasms.
Isolation of Canine Mammary Cells With Stem Cell Properties and Tumour‐Initiating Potential
Authors: Cocola C , Anastasi P , Astigiano S , Piscitelli E , Pelucchi P , Vilardo L , Bertoli G , Beccaglia M , Veronesi MC , Sanzone S , Barbieri O. , Reinbold RA , Luvoni GC , Zucchi I 
Published: July 2009

ContentsRecent data suggest that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self‐renewal or by progenitor cells that have acquired the capacity for cell self‐renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long‐term non‐adherent culture conditions. We showed that cells obtained from spheres that display self‐renewing properties, have multi‐lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour‐initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue.
Oestrogen and Progesterone Receptor Gene Expression in Canine Oocytes and Cumulus Cells Throughout the Oestrous Cycle
Authors: Gonçalves JSA , Vannucchi CI , Braga FC , Paula‐Lopes FF , Milazzotto MP , Assumpção MEOA , Visintin JA 
Published: July 2009

ContentThe aim of this research was to analyze oestrogen receptor‐α (ERα), ERβ and progesterone receptor (PR) gene expression in the canine oocyte and cumulus cells throughout the oestrous cycle. Ovaries from 38 bitches were recovered after ovariohysterectomy and sliced. The phase of the oestrous cycle was determined by vaginal cytology, vaginoscopy and serum hormonal measurements. Oocytes were mechanically denuded by repeated pipetting. For each phase of the cycle, a sample was composed by a pool of 50 oocytes (sample number: prooestrus = 3, oestrus = 8, dioestrus = 5 and anoestrus = 5) or a pool of cumulus cells (prooestrus = 4, oestrus = 7, dioestrus = 4 and anoestrus = 6). Oocyte and cumulus cells’ total RNA was isolated and reverse transcription was conducted to perform real‐time PCR. Oestrogen receptor‐α was expressed throughout the cycle in the oocyte (33.33%, 25.0%, 20.0% and 60.0% for prooestrus, oestrus, dioestrus and anoestrus, respectively) and cumulus cells (50.0%, 47.14%, 25.0% and 66.67% for prooestrus, oestrus, dioestrus and anoestrus, respectively). In the oocyte, the ERβ was also expressed in all phases of the cycle (33.33%, 50.0%, 20.0% and 60.0% for prooestrus, oestrus, dioestrus and anoestrus, respectively), whereas in cumulus cells, ERβ was only expressed during prooestrus (50%) and oestrus (14.29%). Interestingly, while the oocyte PR was not detected in any phase of the cycle, this receptor was expressed during prooestrus (50%), oestrus (42.86%) and anoestrus (16.67%) in cumulus cells. In conclusion, canine oocytes express ERα and ERβ throughout the oestrous cycle, however, there is a lack of PR expression in all these phases. Moreover, in cumulus cells, only ERα was expressed throughout the oestrous cycle.
Expression of the μ Opioid Receptor and Effects of the Opioid Antagonist Naloxone onIn VitroMaturation of Oocytes Recovered from Anoestrous Bitches
Authors: Iorga AI , Valentini L , De Santis T , Ambruosi B , Albrizio M , Guaricci AC , Caira M , Dell’Aquila ME 
Published: July 2009

ContentsThe μ‐opioid receptor (MOR) is expressed in bovine, human, equine and canine oocytes, and in seasonal breeders, it is expressed with higher intensity during the anoestrous phase. Supplementation ofin vitromaturation (IVM) medium with opioid agents, agonists or antagonists, was shown to affect oocyte maturation in several species such as rat, bovine and equine. This study reports the effects of supplementing IVM medium with naloxone (Nx), an opioid antagonist, on nuclear and cytoplasmic maturation rate of oocytes recovered from anoestrous bitches. Cytoplasmic maturation was examined in terms of mitochondrial (mt) distribution. In order to confirm the receptor‐mediated action of Nx, in oocytes of anoestrous bitches, MOR expression was analyzed by Western blot. Cumulus–oocyte complexes, recovered from the ovaries of bitches in anoestrous, were culturedin vitroand Nx was added at the concentrations of 1 × 10−6, 1 × 10−8and 1 × 10−10 m. The rate of oocytes resuming meiosis after culture in presence of 1 × 10−6 mNx (29%) was significantly higher than that of oocytes of control group (12%; p < 0.05). However, treatment with Nx did not affect mt distribution pattern. In denuded oocytes and in corresponding cumulus cells, a doublet of 65 and 50 kDa was observed. We conclude that, in oocytes of anoestrous bitches, MOR is expressed and Nx significantly improves nuclear maturation rate. Further studies should be performed to elucidate the expression of other opioid receptors, such as δ and κ, and possible interactive effects of their antagonists on canine oocyte maturation.
Effects of Short‐term Hyper‐ and Hypoprolactinaemia on Hormones of the Pituitary, Gonad and ‐Thyroid Axis and on Semen Quality in Male Beagles
Authors: Koivisto MB , Eschricht F , Urhausen C , Hoppen HO , Beyerbach M , Oei CHY , Günzel‐Apel AR 
Published: July 2009

ContentsEffects of a short‐term hyper‐ and hypoprolactinaemia on serum concentrations of LH, testosterone and semen quality in six male Beagles were investigated. Blood samples were collected at 3‐day intervals for 12 weeks. The time span was divided into five 3‐week periods: pre‐treatment, metoclopramide (MCP) treatment (0.2 mg/kg orally three times daily), cabergoline (CAB) treatment (5 μg/kg orally once daily), post‐treatment 1 and post‐treatment 2. In the latter, only semen characteristics were evaluated. Semen parameters were analyzed once per week during the whole 15‐week investigation time. At the end of each period, the effects of a single intravenous injection of thyrotropin‐releasing hormone (TRH; 10 μg/kg) on the secretion of prolactin (PRL), LH, testosterone, thyroid‐stimulating hormone and thyroxine (T4) were investigated. Pre‐treatment serum PRL concentration increased under MCP (p < 0.05), followed by a decrease under CAB administration (p < 0.05). Luteinizing hormone and testosterone concentrations were not affected. Except for straight‐line sperm velocity, semen quality did not differ between collection periods. A single iv TRH injection induced a significant PRL increase at 20 min in all experimental periods except during CAB treatment. Luteinizing hormone and testosterone did not show clear TRH‐related changes. Basic T4 levels were significantly reduced after CAB treatment (p < 0.05). The results of the present study demonstrate that MCP‐induced short‐term hyperprolactinaemia in male beagles does not seriously affect the hypothalamo‐pituitary axis and semen quality.
Pregnancy Monitoring in Dogs and Cats Using 3D and 4D Ultrasonography
Authors: Hildebrandt TB , Drews B , Kurz J , Hermes R , Yang S , Göritz F 
Published: July 2009

ContentsThree‐dimensional (3D)/four‐dimensional (4D) volume ultrasound is an established method in human medicine that offers various options for analysing and presenting ultrasound volume data. However, the successful application of the different 3D/4D imaging modalities in pregnant dogs and cats has not yet been reported in the literature. The main reasons for this are: (1) the high costs of 3D/4D ultrasound systems, (2) operation difficulties due to high breathing frequency in non‐sedated animals and (3) the missing specific knowledge in veterinary medicine concerning how to perform high‐quality volume scans. Automatically acquired ultrasound volume data sets were generated with two different ultrasound systems: the portable Voluson i and the stationary Voluson Expert 730. Different 3D/4D imaging modalities were tested in regard of their practicability in pregnancy monitoring in dogs and cats. Nine different volume imaging modalities were applied using the saved files. For the presentation of the static 3D volume data sets, we used the multiplanar, niche, surface, transparency, glass body, inversion, volume calculation and tomographic ultrasound imaging modes. For the dynamic 4D data, the surface and glass body modes were applied. By changing the human standard settings to the requirements of small animal anatomy, it was found that 3D/4D ultrasound has great potential for the characterization of pregnancy in queens and bitches. The 3D/4D technology offered advanced information about pregnancy status and birth prediction and improved the diagnostic confidence. By using standardized examination protocols, 3D/4D ultrasound will allow a reduction in examination time by generating even more relevant information. These benefits, combined with possible future cost reduction of commercial ultrasound systems, might lead to frequent utilization in routine pregnancy diagnostic and birth management in small animal practice.