Triplex doppler ultrasonography to describe the uterine arteries during diestrus and progesterone profile in pregnant and non-pregnant bitches of different sizes.

Hemodynamics of uterine vascularization is modified throughout pregnancy to meet the increasing demand of the growing fetuses and triplex doppler ultrasonography is widely used in human medicine to study the uterine arteries and assess the fetal and placental conditions. The aim of our study was to confirm this observation in the bitch, to evaluate differences between bitches of different sizes and to study abnormal pregnancies. Forty-four bitches were monitored during the estrous period to determine ovulation and every 10 days from ovulation to 50 days post-ovulation: the resistivity (RI) and pulsatility (PI) indexes of the right uterine artery were measured as well as usual assessment of fetal development and follow up of the luteal function. Thirty-three out of forty-four bitches were pregnant, including 6 abnormal pregnancies (resorption of more than 10% of the embryos). We also divided them in four weight categories: 8 were small (<10 kg), 13 medium (10-25 kg), 13 large (>25-40 kg) and 10 were giant breeds (>40 kg). We observed that RI and PI decreased over time and were significantly lower for pregnant bitches compared to non-pregnant ones from 30 days post-ovulation. In contrast, RI and PI did not significantly vary with the size of the bitches and we could not determine a significant impact of abnormal pregnancies either. In conclusion, we found no significant difference related to the size of bitches in the RI and PI. The only significant difference between pregnant and non-pregnant bitches was observed from 30 days post-ovulation.

A review of maternal behaviour in dogs and potential areas for further research.

Here we review information on maternal behaviour in dogs, defined as a combination of all the acts of the mother towards her offspring, which begins before parturition and continues until weaning. In dogs, maternal care is measured using the most commonly observed behaviours, such as the time spent in contact, licking/grooming and nursing of the puppies. Since newborn puppies have a very limited capacity for movement, maternal interaction is essential to their survival, nourishment and protection. It is also an important element of the bonding process between puppies and the bitch and is thought to play a role in the social development of the puppies. Nevertheless, some questions still need to be clarified, such as the best way to quantify factors that may interfere with maternal behaviour. In recent studies, maternal care, or maternal style, was measured using a scoring system and found to be influenced by factors such as litter size, breed and parity, or even human interaction. However, the impact of the emotional state of the bitch and the quality of maternal behaviour on puppy survival and development remain unclear. The long-lasting effects of mother-puppy interactions on puppy behaviour during their adult life are still poorly understood, despite their importance for breeders who wish to prevent future problem behaviours.

The Comet assay for detection of DNA damage in canine sperm.

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno-, terato- or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A-median: 31.55%, interquartile range (IQR): 30.18-38.01; group B-median: 0.90%, IQR: 0.65-1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = -.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non-normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.

A retrospective study of canine prostatic diseases from 2002 to 2009 at the Alfort Veterinary College in France.

A retrospective study was used to investigate the incidence of prostatic diseases in a large population of dogs at Alfort Veterinary Hospital and to clarify epidemiologic features, which might be of a great help to veterinarians in managing and discriminating prostatic disorders. During the investigation period, a total of 72,300 male dogs (coming mainly from the Ile-de-France region) were registered in the Alfort Veterinary College database, and 481 of them (0.7%) were found to have prostatic disorder. The diagnosis was carried out on the basis of clinical signs and ultrasound findings. Among dogs experiencing a prostatic disorder, most frequently recorded diseases were benign prostatic hyperplasia (45.9%) and prostatitis (38.5%), followed by abscesses (7.7%), cysts (5.0%), neoplasia (2.6%), and squamous metaplasia (0.2%). Our study revealed an incidence of 0.3% of prostatic disorders observed in intact male dogs, except in the case of prostatic neoplasia. The mean age of the dogs experiencing prostatic disorders was 8.6 ± 3.2 years. This was significantly different (P < 0.001). Large dogs were significantly more affected by prostatic disorders (P < 0.05), except for prostatic neoplasia. A breed predisposition was suspected in German Shepherd (odds ratio [OR] = 2.1; 95% confidence interval [CI]: 1.5-2.9), Rottweiler (OR = 1.8; 95% CI: 1.2-2.7), American Staffordshire Terrier (OR = 3.8; 95% CI: 2.5-5.8), Berger de Beauce (OR = 3.7; 95% CI: 2.2-6.1), and Bernese Mountain Dog (OR = 2.5; 95% CI: 1.3-4.7).

Fertility in Adult Bitches Previously Treated with a 4.7 mg Subcutaneous Deslorelin Implant.

The absence of fertility problems in male dogs after a single treatment with deslorelin acetate (Suprelorin(®)) is well acknowledged. However, reports on the application of deslorelin in the bitch and information concerning fertility after implant treatment are still limited. In this retrospective study, data concerning induced and spontaneous oestruses of 39 bitches from 17 breeds, treated with deslorelin acetate implants (4.7 mg Suprelorin(®), Virbac, France), were retrieved to assess post-treatment fertility (ovulation rate, pregnancy rate and litter size). Animals were grouped according to treatment characteristics: group 1 (Gr1) - females submitted to oestrus induction, showing natural oestruses afterwards (n = 19); group 2 (Gr2) - females re-implanted with 4.7 mg deslorelin acetate to re-induce oestrus, showing subsequent spontaneous post-implant oestruses (n = 7); and group 3 (Gr3) - females submitted to a 4.7 mg deslorelin acetate implant for oestrus suppression, evaluated at subsequent spontaneous post-implant oestruses (n = 13). Comparison of fertility traits between induced and post-treatment spontaneous oestruses in Gr1 and Gr2 (short treatments), or between spontaneous oestruses after long-treatment schedules (Gr 3) revealed a slightly better performance in spontaneous cycles compared with induced cycles: ovulation rate post-treatment was 97.1%, 94.1% and 94.4% and the pregnancy rate post-treatment was 91.2%, 88.9% and 84.6% for groups 1, 2 and 3, respectively. Nevertheless, fertility in induced and post-treatment oestruses was considered normal. Moreover, the individual litter size did not differ within groups between induced and spontaneous cycles. From these findings, we concluded that treatment with 4.7 mg deslorelin implants did not compromise the bitches' fertility in subsequent oestruses.

Seminal fluid promotes in vitro sperm-oviduct binding in the domestic cat (Felis catus).

From many endangered or threatened species which are expected to profit from assisted reproduction techniques, mainly epididymal sperm of dead or freshly castrated males are available. These sperm had contact to epididymal secretion products but not to seminal fluid components. Notably, products of accessory sex glands have been shown in domestic animals to condition sperm for fertilization, in particular by mediating sperm-oviduct interaction. We report for the first time that motile epididymal sperm from domestic cats are able to bind to fresh oviduct epithelial cell explants from preovulatory females (median [min, max] of 10 [8, 16] and 10 [8, 17] sperm per 0.01 mm(2) explant surface from both isthmic and ampullar regions, respectively). More sperm attach to the explants when epididymal sperm were preincubated for 30 minutes with seminal fluid separated from electroejaculates of mature tomcats (median [min, max] of 17 [13, 25] and 16 [12, 21] sperm per 0.01 mm(2) explant surface from isthmus and ampulla, respectively). The proportion of bound sperm increased from a median of 54% to 62% by seminal fluid treatment. Sperm-oviduct binding could be facilitated by the decelerated sperm motion which was observed in seminal fluid-treated samples or supported by seminal fluid proteins newly attached to the sperm surface. Seminal fluid had no effect on the proportion of sperm with active mitochondria. Extent and pattern of sperm interaction in vitro were independent of explant origin from isthmus or ampulla. Sperm were attached to both cilia and microvilli of the main epithelial cell types present in all explants. In contrast to published sperm-binding studies with porcine and bovine oviduct explants where predominantly the anterior head region of sperm was attached to ciliated cells, the tails of some cat sperm were firmly stuck to the oviduct cell surfaces, whereas the heads were wobbling. Whether this response is a preliminary step toward phagocytosis or a precondition to capacitation and fertilization remains to be determined. In conclusion, treatment of epididymal sperm with seminal fluid or particular protein components should be considered in future investigations for its potential to improve the outcome of artificial insemination in felids.

Presence of membranous vesicles in cat seminal plasma: ultrastructural characteristics, protein profile and enzymatic activity.

This study sought to verify the presence of membranous vesicles in cat seminal plasma by means of transmission electron microscopy and to identify protein profile and some of the enzymatic activities associated with these particles. The transmission electron microscopy observations showed the existence of different sized vesicular membranous structures of more or less spherical shape. These vesicles were surrounded by single-, double- or multiple-layered laminar membranes. The vesicle diameters ranged from 16.3 to 387.4 nm, with a mean of 116.5 ± 70.7 nm. Enzyme activity determinations showed the presence of dipeptilpeptidase IV, aminopeptidase, alkaline and acid phosphatase. To our knowledge, this is the first report that identifies and characterizes the membranous vesicles in cat seminal plasma. However, further studies are necessary to identify the exact site of production of these membranous vesicles in the cat male genital tract and to determine their specific roles in the reproductive events of this species.

Suppression of fertility in adult dogs.

Unfortunately, the overpopulation of dogs is still a problem in the majority of countries and even though surgical methods of sterilization, the most traditional and commonly used technique, have been intensively performed, the impact on the dog population is negligible. The neutering of companion animals as ovariohysterectomy (spaying) or orchidectomy (castration) has its limitations because of the cost, the need of a surgical environment and the risk of surgical and/or anaesthetical complications (ACCD 2009). In fact, surgical castration has been banished in some northern European countries and has limited acceptance in other countries. In a survey performed in Sao Paulo, Brazil, 56.5% of the owners of adopted shelter dogs were against the surgical procedure for different reasons (Soto et al. 2005). Currently, the options for contraception, defined as suppression of fertility are based on hormonal treatment. The treatments can be divided into analogues of gonadotropin releasing hormone (GnRH), progestins and androgens. Other possibilities of contraception are via the immunological system with vaccinations against GnRH, the luteinizing hormone (LH) receptor and the zona pellucida proteins. Finally, there is also the intra-epididymal or intratesticular injection of sclerosing substances in dogs. Mechanical devices to disrupt fertility are not used anymore due to the side effects. Suppression of fertility in adult dogs will be reviewed in order of use and possible impact on the dog population.

Subclinical leishmaniasis associated with infertility and chronic prostatitis in a dog.

A stud dog was presented for acquired infertility. Haematospermia and teratozoospermia were found on two ejaculates 2 weeks apart. A presumptive diagnosis of prostatitis was made follo-wing ultrasound examination. An ultrasound-guided needle core biopsy was performed under general anaesthesia, revealing a mild chronic macrophagic and plasma cell prostatitis with intracytoplasmic amastigotes consistent with Leishmania spp. infection. Presence of Leishmania infantum, Leishmania donovani or Leishmania chagasi was confirmed by polymerase chain reaction in seminal plasma. Serology and serum protein electrophoresis confirmed the diagnosis of a subclinical active systemic leishmaniasis. A meglumine antimoniate and allopurinol treatment was given which clearly improved within 3 months both general condition and the quality of sperm. To the authors' knowledge, this is the first reported case of a prostatitis secondary to a Leishmania spp. infection. Subclinical systemic leishmaniasis should be considered in the differential diagnosis of infertility in dogs suffering from semen alterations.

A real-life study of the use, effectiveness and tolerability of rosiglitazone in France: the AVANCE study.

AIM: The study aimed to determine the effectiveness and tolerability of rosiglitazone, and its profile in terms of treatment adherence, treated patients and prescribing recommendations under everyday conditions of care. METHODS: This was a "real-life" observational longitudinal study including patients with type 2 diabetes mellitus (T2DM) starting treatment with rosiglitazone and followed for up to 2 years. A questionnaire was completed at the time of inclusion and during routine consultations at around 6, 12, 18 and 24 months following inclusion. Information was collected on sociodemographics, clinical history, treatments, co-morbidities, laboratory data and compliance with treatment. There were three primary outcome measures: treatment response (defined as an HbA1c ≤ 8.0% or a decrease in HbA1c ≥ 0.7%); switch to insulin (as considered necessary by the physician); and occurrence of adverse events requiring a change or discontinuation of treatment. RESULTS: The evaluation included 670 patients (61.1%) treated with rosiglitazone/metformin as fixed-dose combination tablets and 427 (38.9%) with standard rosiglitazone tablets. Rates of HbA1c response, defined as an HbA1c less than or equal to 8.0% or a decrease in HbA1c greater than or equal to 0.7%, ranged from 80.6% to 92.1% depending on the follow-up time. The percentage of patients with an HbA1c less than 7% was 18.4% before rosiglitazone was prescribed, and ranged from 48.2% to 57.8% depending on the follow-up period. Sixty-two patients (6.1%, 95% CI: 4.6-7.6%) switched to insulin therapy during the follow-up period. Spontaneously reported adverse events leading to a change or discontinuation of treatment were seen in 45 patients (4.4%, 95% CI: 3.2-5.6%). CONCLUSION: Rosiglitazone showed sustained efficacy, with around 90% of patients defined as responders to the treatment in terms of reduction in HbA1c, and was relatively well tolerated. The adverse-event profile was consistent with the known effects of rosiglitazone, and no signs of increased cardiovascular ischaemic risk were observed. These results are in agreement with previous studies on rosiglitazone.

Folliculogenesis, ovulation and endocrine control of oocytes and embryos in the dog.

Reproductive physiology in dogs is quite unusual compared with that in other mammalian species. The peculiarities include the presence of numerous polyoocyte follicles, the ovulation of an immature oocyte (GV stage, non-fertilizable) and a peri-ovulatory period during which concentrations of circulating progesterone are particularly high. The aim of this review is to examine the unusual aspects of the reproductive physiology of dogs and how this relates to the clinical biology of this species.

Results of GnRH agonist implants in oestrous induction and oestrous suppression in bitches and queens.

Over the last 10-15 years, long-acting GnRH agonists have become widely available. In the field of small animal reproduction, most recent studies have focused on the use of two compounds developed under the form of subcutaneous implants: azagly-nafarelin and deslorelin. Only the latter has been commercially available for use in male dogs, first in Australia and New Zealand, then in several countries of the European Union since 2008. Although officially marketed for male dogs, this compound has also been studied in bitches and more recently in queens. Some published papers or recent presentations at congresses--still unpublished--have focused on the use of GnRH agonists implants in females.

Induction of fertile oestrus in the bitch using Deslorelin, a GnRH agonist.

Oestrus induction in various canine breeds was attempted in 32 bitches. A group of 8 bitches were treated 80-160 d following their previous oestrus (G1) whereas a second group of 24 bitches (G2) were implanted 200-590 d following their previous oestrus. The treatment for each bitch consisted in one Deslorelin implant (Suprelorin® 4,7 mg, Virbac, France), inserted subcutaneously in the post-umbilical region. Ovulation, pregnancy rate and litter size were recorded. All bitches came in heat 4.3 ± 1.4 d after implantation (2-7 d). Ovulation was reported in 62.5% in G1 and 87.5% in G2. One bitch refused mating and since no AI was performed, she was not considered for further analysis. Pregnancy was obtained in 25% in G1 versus 78.3% in G2. Mean litter size was 6.7 ± 3.5 puppies (1-14). Luteal failure was suspected in 3 bitches, two that remained non-pregnant and one which aborted 58 d post-ovulation since the owner refused progesterone supplementation. Deslorelin implants can therefore be considered as a valuable alternative to induce fertile oestrus in bitches in anoestrus. Follow-up of the luteal phase is recommended, since some bitches might encounter luteal failure.

Estimated pregnancy length from ovulation to parturition in the bitch and its influencing factors: a retrospective study in 162 pregnancies.

An accurate timing of parturition is very useful for managing canine parturition. It is generally accepted that parturition in bitches occurs between 64 and 66 days after the luteinizing hormone peak. In this retrospective study, we determined pregnancy length in different breeds and its influencing factors dating it from the estimated day of ovulation (EDO), defined as the day when peripheral plasma level of progesterone (P4) reaches 6 ng/ml. From January 2001 to December 2006, 162 pregnancies in 151 bitches of 53 different breeds were followed. Different parameters concerning the bitch, the litter, the type of semen and the type of reproduction were studied. The mean estimated pregnancy length in the bitch from EDO to parturition was 63.1±2.1 days. The main influencing factors for the pregnancies studied were the breed, the size of the bitch and the number of puppies within the litter.

Clinical use of GnRH agonists in canine and feline species.

GnRH (gonadotrophin releasing hormone) is a key hormone of reproductive function in mammals; agonist forms have been largely developed, and data concerning their use in small animal reproduction are now abundant. GnRH agonists act by a two-step mechanism. First, their agonist properties on the pituitary will cause marked LH (luteinizing hormone) and FSH (follicle-stimulating hormone) secretion into the bloodstream, accompanied by an increase in the concentrations of sex steroid hormones. Then, in case of constant administration, GnRH agonists will lead to pituitary desensitization, and FSH and LH levels will collapse. These two effects have been widely documented, and these compounds have many potential benefits in a clinical context, capitalizing both on their stimulating and sterilizing effects.

Anthropometric characteristics and cardiometabolic risk factors in a sample of urban-dwelling adults in Senegal.

AIM: The aim of this study was to analyze the relationship between anthropometric characteristics and cardiometabolic risk factors in urban-dwelling adults in Senegal to evaluate future threats to the public health in terms of chronic diseases. METHODS: Age- and gender-matched control subjects for a study on the prevalence of lipodystrophy in HIV+ patients were selected between June and September 2006 from the general population through systematic home visits guided by area of residence of cases. After consenting to participate, these subjects underwent anthropometric, clinical and biological examinations in their homes. RESULTS: The sample included 60 men and 106 women, mean age of 43.2 ± 9.4 years. Although the prevalence of overweight and obesity was much higher in women (30.2 and 29.2%, respectively) vs. 23.3 and 3.4%, respectively, in men (P<0.001), the women had lower waist-to-hip ratios (mean [95% CI]: 0.78 [0.77-0.80] vs. 0.86 [0.84-0.88] in men; P<10(-4)) and better systolic blood pressure, triglyceride and high-density lipoprotein (HDL)-cholesterol levels. However, their insulin levels were significantly higher (32.1 [28.2-36.5] pmol/l vs. 25.5 [21.0-30.8] in men; P<0.04). Principal component analysis showed that glucose and insulin correlated with subcutaneous fat, whereas blood pressure correlated with central fat distribution. Lipids were distributed between these two factors. CONCLUSION: Obesity still appears to be rare in Senegalese urban-dwelling men, whereas women, despite their overweight, have no untoward cardiometabolic profiles. However, the observed correlations between cardiometabolic risk factors and the amount and/or distribution of body fat suggest that obesity prevention should not be overlooked in the public health agenda for sub-Saharan Africa.

Effect of post-thaw dilution with caffeine, pentoxifylline, 2'-deoxyadenosine and prostatic fluid on motility of frozen-thawed dog semen.

The aim of the experiment was to evaluate the motility pattern of frozen-thawed canine semen to which pentoxifyilline (PTX), caffeine (CAF), 2'-deoxyadenosine (DX), and prostatic fluid (PROST) were added after thawing. Semen evaluations were performed using computer-assisted sperm analysis (CASA) at thawing and during 120min of incubation at 37 degrees C. Three experiments were conducted: 1) to establish which concentrations of stimulants work best; 2) to investigate the interaction between thawing rate and addition of CAF 5mM, PTX 2.5mM and PROST; 3) to evaluate the effect of PTX 7.5mM and DX 5mM on semen motility after thawing. In experiment 1, ALH and VCL were enhanced at thawing by CAF 7.5mM (CAF 7.5: 9.1+/-0.5microm; control: 6.7+/-0.4microm) and DX 5 and 7.5mM (DX 5: 199.1+/-12.8microm/s; DX 7.5: 197.3+/-13.9microm/s; control: 162.5+/-8.4microm/s), while PTX 2.5-5-7.5mM improved TOT after 120min of incubation. In experiment 2, PROST lowered ALH values throughout incubation (P<0.05) with respect to the other treatments, in particular when compared to CAF at Time=30 and at Time=60. In experiment 3, PTX 7.5mM improved VAP (PTX: 101.6+/-6.8microm/s; control: 81.9+/-10.5microm/s), VSL (PTX: 82.9+/-6.4microm/s; control: 65.9+/-9.8microm/s), VCL (PTX: 214.3+/-13.3microm/s; control:167+/-15.7microm/s), ALH (PTX: 10.5+/-0.3; control: 7.3+/-1.4microm), PM (PTX: 11.3+/-4.2%; control: 7.7+/-3.9%) and TOT (PTX: 20.1+/-5.3%; control:15.6+/-5.6%) at Time=120, while DX 5mM influenced VCL at Time=60 (DX: 218.3+/-14.3microm/s; control: 188.5+/-7.5microm/s, P<0.05). Motility stimulants may be useful for enhancing motility of canine frozen-thawed spermatozoa without affecting sperm longevity.

Induction of parturition with aglepristone in various sized bitches of different breeds.

The objective of this study was to confirm in various breeds of dogs the efficacy and safety of a parturition induction treatment described to be successful in Beagle dogs. Parturition was induced in seven various sized pregnant bitches of different breeds, with 15 mg aglepristone per kg at day 59-61 post-estimated ovulation day, followed 24 h later by 0.15 IU oxytocin per kg subcutaneous injections every 2 h. Two bitches were small-sized bitches (<10 kg), three bitches were large-sized bitches (30-40 kg) and two bitches were giant bitches (>40 kg). The results were compared to a control group (n = 6), in which bitches underwent a natural delivery in the same environmental conditions as the induced group. In the induced group, parturition was successfully induced in 7/7 bitches. The first pup in a litter was born on average 25.9 +/- 3.29 h after aglepristone administration (21-30 h). Two of seven bitches from the small-sized group delivered some of their pups before the first administration of oxytocin. The mean duration of parturition was 9.6 +/- 5.4 h vs 8.0 +/- 4.8 h in the control group. The mean interval between two successive pups being delivered was 115.6 +/- 82.8 min (34-265) vs 68.8 +/- 24.5 min in the control group (p < 0.03). The mean weight at parturition did not differ significantly between the two groups. One litter of four Yorkshire Terrier pups in the induced group were premature at the time of birth and died between 19 and 29 h post-delivery. This study, although on a very limited number of dogs, confirms the efficacy of the aglepristone/oxytocin protocol to induce parturition in dogs.

Elective caesarean operation in the bitch using aglepristone before the pre-partum decline in peripheral progesterone concentration.

The aim of this study was to investigate caesarean operation (CO) undertaken before the pre-partum decrease of progesterone but following administration of a progesterone receptor antagonist and to evaluate the innocuity of this procedure for the dam and pups. Thirty seven bitches of 15 different breeds, received an injection of 15 mg/kg aglepristone 59 or 60 days after the estimated day of ovulation, determined by progesterone quantitative assays, and caesarean section (CS) was performed between 20 and 24 h after administration. Progesterone remained above 6 nmol/l at the time of CS (mean = 15.75, SD = 3.84). No post-operative clinical complications were reported in any of the bitches. All bitches were able to nurse and feed their puppies in the first 24 h following surgery. No pups showed any signs of prematurity and 5 out of 188 pups (2.6) died in the first 2 weeks after delivery. This small study demonstrates that a CS may be safely and successfully performed an average of 2 days before the expected date of parturition following the administration of aglepristone, without any harmful consequence for the dam and her neonates.