Analysis of biochemical parameters in canine fetal fluids during the second half of pregnancy.

Although fetal fluids, amnion (AM) and allantois (AL), have been studied at the end of pregnancy in the bitch, little is known about their composition during gestation. The aim of this study was to characterize and compare the biochemical parameters of fetal fluids collected from healthy pregnant bitches in mid pregnancy. Fetal fluids were collected from 26 pregnant bitches between 35 and 62 days from ovulation based on ultrasonography. A total of 149 samples were analyzed; 94 samples of known origin (61 a.m. and 33 AL) from 17 dams, which included 17 paired samples (AM and AL from the same fetus; 34 samples), and 55 of unknown origin from nine dams. Reference ranges were formulated for 22 biochemical parameters in AM and AL based on the samples of known origin (n = 94). All parameters (creatinine, albumin, GGT and urea) significantly different between AM and AL were included (by mixed model ANOVA) in the analysis and an algorithm to predict the origin of the fluid was developed (by logistic regression). Overall, of the 94 samples, 79 samples (84%) were correctly identified by the algorithm. For the 33 allantois samples, and for the 61 amnion samples, 23 (69.7%) and 56 (91.8%) were correctly identified by the algorithm, respectively. Based on the algorithm, 31 of the unknown samples (58%) were classified as amnion and 22 samples (42%), as allantois fluid. For all the unknown samples, the average certainty was 86% (compared to 81% for the samples whose fluid type was known), showing the degree of precision of the classification.

Short communication: Progressive motility of frozen-thawed canine semen is highest five minutes after thawing.

Progressive motility is usually estimated by visual inspection using a light contrast microscope at X 100 immediately after semen collection or immediately after thawing frozen semen. Standard operating procedures have never been established for this test. The objective of this experiment was to examine time-dependent changes of motility after thawing cryopreserved canine semen. Semen of 35 dogs was collected, and volume, concentration, progressive motility, morphology, membrane integrity and HOS test were evaluated. For cryopreservation, CaniPRO® Freeze A&B was used. Semen was thawed and diluted using CaniPRO® culture medium. After thawing, semen was evaluated as before. In addition, every sample was evaluated for progressively motile sperm cells 0, 5, 20 and 60 min after thawing. Progressive semen motility was significantly highest five minutes after thawing.

Cystic ovaries and ovarian neoplasia in the female dog - a systematic review.

Most common ovarian diseases appear to be cystic or ovarian tumours. These conditions may affect health, fertility or even the life of the animal. Literature related to ovarian cysts and neoplasia in dogs was assessed to summarize studies and case series on ovarian cysts and primary ovarian tumours in regard to epidemiologic, clinical and fertility aspects. Two research papers on histopathological examination of ovarian cysts could be identified. Seven research papers on ovarian tumours were also assessed, showing inconsistent results on the frequency of occurrence of different types of neoplasia. This might be due to different definitions of diseases and different diagnostic procedures. In conclusion, scarce research has been performed on ovarian diseases in the bitch in the last decades. Many publications are based on case reports. Most quality criteria of study methodology like blinding, randomization, sensitivity and specificity of test methods are not considered in papers on ovarian diseases. In the future, a multicentre data collection of cases could help to gather more data on the incidence, clinical relevance, treatment options and prognosis of ovarian diseases.

Prognostic value of a pre-freeze hypo-osmotic swelling test on the post-thaw quality of dog semen.

Throughout cryopreservation, sperm are exposed to major osmotic challenges. Only intact membranes of sperm cells are able to regulate these volumetric changes, which can be determined by the hypo-osmotic swelling test (HOS test). Correlations between the HOS test and conventional semen variables are inconsistent. Therefore, the objectives of this study were (1) to examine relationships between HOS test results and standard semen variables before freezing and after thawing and (2) to evaluate the prognostic value of the HOS assessments on post-thaw quality of dog semen. Semen of 35 dogs was collected and analyzed before freezing and after thawing following a 7-day freeze-thaw interval. Conventional semen variables such as sperm cell motility, membrane integrity morphology were evaluated and the HOS test was conducted with results from this test being recorded. In fresh semen the HOS test was positively correlated with progressive motility of sperm cells: r=0.52, sperm cell membrane integrity: r=0.50 and normal sperm cell morphology: r=0.46 (P<0.05). In frozen-thawed semen, the data obtained with the HOS test were positively correlated with progressive sperm cell motility: r=0.67 and membrane integrity: r=0.86 (P<0.05). The data obtained with the HOS test in fresh semen were positively correlated with sperm cell membrane integrity: r=0.50 normal sperm cell morphology: r=0.55 and data from the HOS test (r=0.43; P<0.05) with frozen-thawed semen. For the prediction of individual cryopreservation capacity, results from assessment of the fresh semen variables of good and poor semen quality were statistically compared. Based on these results, it is not possible to predict the quality of frozen-thawed dog semen using the HOS test.

Evidence-based Medicine in Animal Reproduction.

With new knowledge being generated and published daily, the importance of evidence-based approaches in veterinary medicine is obvious. Clinicians must stay current or risk making poor decisions that clients may challenge. Especially in animal reproduction, several new substances and procedures to diagnose or treat reproductive disorders have been introduced in the last years. On the other hand, a closer look at the quality of published literature on animal reproduction reveals major deficits in methodology and reporting of many clinical trials. We strongly recommend systematically assessing the quality of scientific information when reading journal papers before using the given information in practice. The aim of evidence-based medicine (EBM) is to base the decisions in the practice of medicine on valid, clinically relevant research data. Therefore, we suggest that students should become familiar with the concepts of evidence-based veterinary medicine (EBVM) at the beginning of their veterinary education. Concepts and supporting tools such as checklists for literature assessment have been developed and validated. The purpose of this article is to review and discuss the importance of incorporating EBVM in animal reproduction. The need for further research that produces strong evidence in different fields of animal reproduction and better reporting of relevant study information is obvious.

Does oral administration of the amino acid tyrosine affect oestradiol-17ß concentration and sexual behaviour in the bitch?

The oral administration of the amino acid, tyrosine, has been for years recommended in order to improve fertility, especially to improve copulation behaviour in female dogs. However, evidence is comparatively poor. The objective of our study was to determine whether oral administration of tyrosine has an effect on oestradiol-17ß concentrations and the oestrous behaviour in the bitch. Fifty bitches were randomly allocated to one of two treatment groups in which each dog received 100mg/kg/day of either tyrosine or milk sugar orally between Day 3 and Day 9 of heat. Every two to three days, a gynaecological examination was performed and blood samples were taken to determine oestradiol-17ß and progesterone concentrations. The day of ovulation was estimated by clinical findings, and according to the specifications of the laboratory, once progesterone values exceeded 12.7nmol/l. The observed copulation behaviour was not different between the groups. No differences in volume and visual nature of vaginal discharge were observed. At the day of ovulation, mean oestradiol-17ß concentration in the treated group was 163.4pmol/l and 162.2pmol/l in the placebo group, respectively. In conclusion, feeding tyrosine to female dogs between Day 3 and Day 9 of heat did not alter visual signs of heat or copulation behaviour, and did not alter oestradiol-17ß concentration.

Endoscopic resection of a vaginal septum in a bitch and observation of septa in two related bitches.

CASE HISTORY: In 2008, a 3-year-old female Labrador Retriever was presented for routine breeding soundness evaluation. CLINICAL FINDINGS: Vaginal endoscopy revealed an 8-cm long and 1-cm thick vaginal septum. Surgical resection of the septum was performed under general anaesthesia using a bipolar high-frequency resectoscope. Three months later the bitch was naturally mated and subsequently delivered five male and two female puppies without complications. Both female puppies were examined at the age of 12 and 15 months and showed a vaginal septum with a diameter of 0.3 and 0.5 cm, respectively. DIAGNOSIS: Vaginal septa in three related bitches. CLINICAL RELEVANCE: This is the first report describing the use of a bipolar high-frequency resectoscope for vaginal surgery in dogs. The septum could be resected quickly, with preservation of surrounding structures. Furthermore, it is the first report collecting information about familial presentation of vaginal septa in female dogs. Based on this case, we suggest that consideration should be given to the possible risk of inheritance of vaginal septa before considering surgical resection and breeding of female dogs.

Follicular cysts and prolonged oestrus in a female dog after administration of a deslorelin implant.

CASE HISTORY: A 7-year-old female Bernese Mountain dog was presented for treatment to prevent oestrus. One 4.7-mg deslorelin implant was injected S/C between the shoulder blades. Three months later, the dog was re-examined because of prolonged oestrus. CLINICAL FINDINGS: Ultrasound showed multiple thin-walled cysts on both ovaries. Blood samples were taken at various intervals during the following weeks. Concentrations of oestradiol and vaginal smears indicated pro-oestrus. Follicle stimulating hormone (FSH) and luteinising hormone (LH) remained at basal concentrations. DIAGNOSIS: Follicular cysts and prolonged oestrus. TREATMENT: Two treatments of 360 IU human chorionic gonadotrophin (hCG) were administered 42 days apart, that induced a partial or complete degeneration of the cysts. However, concentrations of oestradiol rose again within a few weeks. After the second treatment with hCG the bitch developed pyometra. After successful conservative treatment with antibiotics and aglepristone the dog underwent ovariohysterectomy. CLINICAL RELEVANCE: Slow-release biocompatible implants containing deslorelin provide an opportunity for effective contraception for male dogs. Some authors describe deslorelin also as safe and efficacious for the use in female dogs but those recommendations are derived from few studies, with a small number of animals. Based on the outcome of this case and the experience of other authors we suggest a complete gynaecological examination be performed before implanting deslorelin, and weighing the risks against the advantages in controlling fertility. It is well known that the risk of induction of oestrus can be reduced when implants are administered at concentrations of progesterone in plasma of ≥16.0 pmol/L. The implant should be administered S/C, cranial and close to the umbilicus, to have the option of relocation and excision if necessary.