Insights into alternative cryoprotectants to freeze sperm of domestic cats.

Globalization and habitat destruction pose a significant threat to wildlife felids. Even though conservation banks for genetic materials have been created, the sperm cryopreservation with minimal cell damage is still a great challenge. Thus, this study aimed to compare the effects of two commercial extenders with different concentrations of alternative cryoprotectants on thawed sperm quality of domestic cats. Five adult cats were anaesthetized (using a combination of 40 µg/kg medetomidine associated to 5 mg/kg ketamine), and the semen was collected by electroejaculation (electrical stimulation of 2-3 V). Semen samples were evaluated for sperm characteristics (kinetics, morphology, membrane integrity and morphometry). Subsequently, they were sorted into two aliquots and centrifuged. The aliquots were added to a commercial extender containing 3% glycerol and 2% methylformamide (extender I) or 2% glycerol and 3% methylformamide (extender II), frozen, thawed (37°C/30 s) and reevaluated. Comparatively, the sperm kinetics and membrane integrity of fresh semen were higher (p < .002) than frozen samples in extender I and II. Total and progressive motility were lowest in the thawed samples. However, the subjective analysis indicated high sperm motility, since the kinetics evaluation was impaired by the low cell number in the thawed samples. There were no differences in sperm morphology between the groups. In the sperm morphometric analysis, a significant difference (p = .04) was identified in the length of the intermediate piece in extender II samples compared with fresh and extender I. Thus, it can be concluded that although the concentrations tested did not maintain the kinetic parameters and membrane integrity of spermatozoa after thawing, the extender with a lower concentration of glycerol was less toxic for maintaining the midpiece length.

The emergence of Brucella canis as a public health threat in Europe: what we know and what we need to learn.

The zoonotic bacteria, Brucella canis, is becoming the leading cause of canine brucellosis in Europe. In dogs, it causes reproductive problems as well as non-specific lameness or discospondilitis. In humans, B. canis can be origin of chronic debilitating conditions characteristic to its genus such as undulant fever, splenomegaly, and lymphadenopathy. Although B. canis shows some pathogenic characteristics similar to B. abortus and B. melitensis, it lacks surface O-polysaccharide, like nonzoonotic B. ovis. This review shows that host-B. canis interactions are still poorly understood, with many knowledge and capability gaps, causing relatively poor sensitivity and specificity of existing diagnostic tools. Currently, there is no vaccine for this rough Brucella species. Besides, antimicrobial therapy does not guarantee bacterial elimination, and infection relapses are frequently reported, increasing the risks of antibiotic resistance development. B. canis has been detected in dogs in almost all European countries which increased human exposure, but currently there is no systematic surveillance. Moreover, B. canis caused brucellosis is not included in Animal Health Law, and therefore there is no legal framework to tackle this emerging infectious disease. To map out the diagnostic strategies, identify risks for human infections and propose management scheme for infected pet and kennel dogs, we present current understanding of canine B. canis caused brucellosis, outline major knowledge gaps and propose future steps. To address and highlight challenges veterinary and public health services encounter in Europe, we developed two B. canis infection scenarios: of a single household pet and of a kennel dog in larger group.

Effect of male age on semen quality in domestic animals: potential for advanced functional and translational research?

Age and other factors like season and breed are often associated with sperm quality and fertility in domestic animals. Even though many studies assessed the relationship between the age of the male and sperm parameters, the effects have not been comprehensively evaluated. Changes in semen quality from pubertal (young) to adult and old age were identified in the bull, ram, buck, boar, dog, and stallion, respectively. The review discusses the association between male age and semen volume, the total number of spermatozoa per ejaculate, sperm concentration, motility, morphology, sperm cell function, sperm DNA integrity, oxidative stress, and antioxidant activity in these species of animals. Generally, semen characteristics improve to a certain age, which declines as the animal ages. Only a few studies evaluated the impact of advanced age or employed advanced functional sperm assessment methods to assess age-related changes in sperm quality and male fertility. Such studies in the dog or stallion, for instance, may contribute to advancing knowledge in human-assisted reproductive techniques used in patients of advanced paternal and maternal age.

Causes of pregnancy arrest in the canine species.

In the canine species, early pregnancy arrest before 30-40 days of pregnancy will induce intra-uterine embryonic or foetal resorption, with very few clinical signs. If no genital examination by ultrasound is performed at that time, it will often remain unnoticed and the bitch will be qualified as infertile. It is only when pregnancy stops at a later stage, mostly after 40 days, that clinical signs will be seen. Expulsion of aborted foetuses or placentas may be observed, although the dam frequently eats the expelled tissues. Intra-uterine mummification may also occur. This article reviews the causes of pregnancy termination in the bitch, both at the embryonic and the foetal stages, reported in the literature. Canine brucellosis is by far the most important disease concerned in this aspect. There is an important current concern about this disease, due to the fact that several outbreaks have been observed recently in Europe, and that it is a very contagious disease; it may be an underestimated zoonosis. Other bacterial causes of pregnancy arrest are sporadic. There is, however, a growing attention towards the microbiological content of raw food diets which are more and more popular among dog breeders and may-if badly prepared-contain abortifacient bacteria such as Campylobacter jejuni or Listeria monocytogenes. The abortifacient role of endogenous vaginal bacteria and mycoplasms remains unclear and may be related to an imbalanced vaginal flora with subsequent ascending bacterial contamination of the uterus. The role of Canine Herpesvirus on abortion is controversial and probably not frequent. Other viruses have been shown experimentally to induce abortion but their natural occurrence in this respect remains unknown. The same applies to the parasite Neospora caninum which is suspected, but not proven, potentially to induce pregnancy arrest in bitches. Among non-infectious causes, uterine pathology such as cystic endometrial hyperplasia (CEH) or sub-clinical post-mating endometritis which can cause infertility and may also induce embryonic resorption. The role of luteal insufficiency in pregnancy arrest is probably overestimated.

Uterine issues in infertile queens: Nine cases.

This prospective case series investigated potential uterine causes of infertility in queens. Purebred queens with infertility (failure to conceive, embryonic death, or failure to maintain pregnancy and produce viable kittens), but no other reproductive disorders were examined approximately 1-8 weeks before mating (Visit 1), 21 days after mating (Visit 2), and 45 days after mating (Visit 3) if pregnant at Visit 2. Investigations included vaginal cytology and bacteriology, urine bacteriology, and ultrasonography. At Visit 2 or 3, uterine biopsy or ovariohysterectomy was performed for histology. Of nine eligible queens, seven were non-pregnant by ultrasound at Visit 2 and two had lost pregnancies by Visit 3. Ovulation was confirmed by serum progesterone concentration in all queens. Ultrasonic appearance of the ovaries and uterus was compatible with a healthy status except for one queen with signs of cystic endometrial hyperplasia (CEH) and pyometra, a follicular cyst in another, and fetal resorptions in two queens. Six cats had histologic lesions of endometrial hyperplasia, including CEH (n=1). Only one cat had no histologic uterine lesions. Bacteria were cultured from vaginal samples in seven queens at Visit 1, (two were non-evaluable), and in five of seven queens sampled at Visit 2. Uterine cultures were negative except for the cat with pyometra. All urine cultures were negative. In summary, the most frequent pathology observed in these infertile queens was histologic endometrial hyperplasia, which can potentially inhibit embryo implantation and healthy placental development. This suggests that uterine disease might contribute substantially to infertility in purebred queens.

Infertility In Queens: Clinical approach, experiences and challenges.

CLINICAL CHALLENGES: It is not easy for a veterinarian to determine the cause of infertility in a queen. The simplest method is to start by investigating the most common causes and then progress by successive elimination to the less common causes. For example, mating problems are commonly responsible for reproductive failure, and owners should be particularly vigilant in checking that the mating is progressing satisfactorily. Frequently, a veterinarian may have to determine, among other things, if there is a uterine or hormonal disorder or if an infectious pathology may be involved. In the case of a queen that is not cycling, it is important to differentiate a lack of onset of puberty in a young queen from a lack of cyclicity in an adult queen. AIM: This review sets out a practical, step-by-step approach that veterinarians can follow, starting with the most common causes of infertility in queens and moving towards more rare or specific causes. EQUIPMENT AND TECHNICAL SKILLS: Simple techniques such as vaginal smears are essential to determine if a queen that is ready to be mated is in full oestrus, if a queen is cycling or not and to diagnose inflammation of the genital tract. Veterinarians should be familiar with the peculiarities of the swabbing technique in cats, as well as the interpretation of feline vaginal cytology. Progesterone assays are useful on many occasions in the diagnosis pathway. A laboratory capable of performing karyotypes should ideally be available. Finally, it is important to be able to perform an ultrasound examination of the ovaries and uterus in a queen. EVIDENCE BASE: Although it appears that infertility is a problem among purebred queens, there is a lack of knowledge in this area and too few published studies. Some causes of feline infertility remain poorly investigated.

Hypo-Osmotic Swelling Test (HOST) for Feline Spermatozoa: The Simplified Procedure and the Aspect of Sperm Morphology.

Hypo-osmotic swelling test (HOST) is used to assess the functional integrity of sperm plasma membranes in many species. The primary aim of this study was to test a simplified HOST procedure for the evaluation of feline semen. The second objective was to check if sperm abnormalities can influence the results of this test. Urethral semen was collected from 19 male, domestic cats. In Exp. 1, HOST was performed in different media (50 mOsm/kg fructose or distilled water), temperature (37 °C or room temperature) and time (5 and 30 min). In Exp. 2, the potential effect of sperm abnormalities on HOST results was assessed by observing individual normal and abnormal spermatozoa microinjected into droplets of distilled water. The results showed no differences between the HOST results performed in different media, temperature and time. Viable abnormal spermatozoa were able to swell under hypo-osmotic conditions in the same manner as normal ones, except spermatozoa with distal droplets, which showed a higher frequency of 'despiralization'. In conclusion, HOST can be reliably performed at 0 mOsm/kg for 5 min at room temperature, which may contribute to a wider use of this test under clinical environments. Viable abnormal spermatozoa are able to swell under hypo-osmotic conditions; therefore, their presence in the ejaculate would not bias the results of HOST when total coiling is calculated.

Profile of Dogs' Breeders and Their Considerations on Female Reproduction, Maternal Care and the Peripartum Stress-An International Survey.

Dogs are part of many households worldwide and in recent years in some western countries, more and more people describe them as members of the family. The accurate number of dogs per household and the actual demand for puppies are hard to estimate correctly. The sources of dogs seem to be an important element to consider before acquiring a dog since some behavioural disorders are more likely observed in dogs coming from puppy mills and pet stores. Therefore, there is an increased need to better-know dog breeders, theirs believes and practices. To full-fill this objective, an online questionnaire in five different languages (English, German, Polish, Portuguese and Spanish) was sent to dog breeders. The survey intended to study the demographics of dog breeders and to understand their perception and practices about reproduction, maternal behaviour and management of the dam during the peripartum period. We were also interested to evaluate the occurrence and the impact of stress around parturition and how breeders deal with overstressed bitches and if they believe that motherly quality could have a later-on effect on the livelihood of the dog. Six hundred and sixty-eight respondents participated in the survey, mainly from Australia, Brazil, Canada, Germany, Portugal, Poland, Spain, United Kingdom, United States of America and non-listed country (reported as 'other' in the survey). A country effect was observed in relation to housing system, human interaction, the use of techniques to define breeding and whelping time and different methods to manage stress during the peripartum period. Considering the demographics of the responders, breeding activity is, in general, a family based activity with an effect of country. Use of progesterone levels to manage reproduction and pregnancy diagnosis by ultrasound were cited as common practice. In general, parturition takes place under human supervision. Peripartum period was perceived as a stressful moment with a difference in the way of addressing the problem based on the country but reassuring the bitch by increasing human presence was frequently described. Inappropriate maternal behaviour was observed by half of responders and if presented primiparous bitches and parturition by C-section were overrepresented. Puppies stayed with the dams from 4 weeks to 9 weeks and dog breeders from Spain, Poland and Portugal are keeping puppies with their mother the longest. Overall, poor maternal behaviour has an impact on puppies' cognitive development with German breeders being more convinced than their counterparts from Brazil and Spain.

Evaluation of surface plasmon field-enhanced fluorescence spectroscopy for rapid measurement of progesterone concentration in bitches.

OBJECTIVE: To compare progesterone (P4) concentrations measured with surface plasmon field-enhanced fluorescence spectroscopy (SPFS) and chemiluminescence immunoassay (CLIA) in serum and plasma samples of client-owned bitches of various ages and breeds and to determine reference ranges for P4 concentrations at various stages of the estrous cycle. SAMPLES: 102 serum samples and 104 plasma samples. PROCEDURES: In experiment 1, 1 aliquot each of serum and plasma was analyzed for P4 concentration by use of SPFS incorporated in a veterinary-specific point-of-care immunologic analyzer and CLIA. In experiment 2, serum collected from bitches in various stages of the estrous cycle was analyzed for P4 concentration by use of SPFS to establish reference ranges for each stage. RESULTS: In experiment 1, P4 concentrations measured by SPFS and CLIA were highly correlated (serum, r = 0.966; plasma, r = 0.968). In experiment 2, ranges of serum basal (proestrous) P4 concentrations (n = 114) and P4 concentrations at the estimated time of ovulation (76), during pregnancy or diestrus (107), and during the prepartum period (50) measured with SPFS were 0.42 to 1.46 ng/mL, 3.69 to 7.85 ng/mL, 11.73 to 28.24 ng/mL, and 1.54 to 3.22 ng/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Because serum and plasma P4 concentrations measured with SPFS were highly correlated with those measured with CLIA and ranges of serum P4 concentrations measured with SPFS for each of phase of the estrous cycle were well-defined for the large sample size, veterinarians may be able to accurately use this veterinary-specific point-of-care immunologic analyzer with SPFS methodology to determine P4 concentrations of bitches in their daily practice.

Influence of ADAPTIL® during the Weaning Period: A Double-Blinded Randomised Clinical Trial.

ADAPTIL®, a dog-appeasing pheromone, was shown to modify the dam-puppies' interactions during the neonatal period but could also influence the weaning period. Fourteen bitch/litter dyads continuously exposed to ADAPTIL® from the third/fourth weeks until the seventh/eighth weeks postpartum were compared to 11 dyads exposed to a placebo. Maternal and puppy behaviours were video-recorded, and at three time points (weeks three/four, weeks five/six and weeks seven/eight) after parturition. The well-being of the puppies and the overall relationship with the bitch were assessed using visual analogue scales (VAS) completed by the caregivers. All mothering behaviours, such as time of contact, licking and the amount of time dedicated to nursing puppies, decreased gradually from weeks three/four to weeks seven/eight. A switch in nursing position was observed over time: the use of the standing position increased compared to the lying position. The treatment had an effect on the nursing position: bitches in the ADAPTIL® group nursed more often in a lying (p = 0.007) or sitting position (p = 0.037), whereas for the placebo group, they favoured the standing position (p = 0.011). Once the puppies became more demanding for suckling, the bitches started showing rejection signs or aggressive growling, with a peak at weeks seven/eight. The pheromone seemed to reduce the intensity of avoidance in bitches exposed to ADAPTIL® at all time points. The score of all events combined as a sign of frustration showed a difference over the full period (p = 0.003), with the placebo group having a significantly higher score. From the caregiver perspective (through the VAS), under ADAPTIL®, the bitches were calmer when puppies tried to suckle (p = 0.001), more tolerant towards pups (p = 0.025), showed a greater motherly attitude (p = 0.016), the puppies cried less when left alone (p < 0.001) and interactions amongst pups were more harmonious (p = 0.055). Under ADAPTIL®, the bitches were less annoyed by the puppies, who seemed to cope better with frustration. The breeders perceived a benefit of the pheromone during the weaning period.

Infertility in purebred cats - A review of the potential causes.

Although purebred cat breeding is growing in popularity in European countries, most research and publications concern dog reproduction. Infertility in queens has been poorly studied. It may not come as a surprise as common domestic shorthair cats are well known for their excellent fertility. However, even in the latter, the infertility rate is around 20%. Only recently, published articles have analyzed the reproductive performances in different breeds, for example in UK, Sweden, France and Italy, and found similar figures. In cats, infertility may arise from many factors such as mismating, prolonged anestrus or silent heats, hormonal, nutritional, genetical or chromosomal causes. Also, infectious diseases, probably frequent in breeding facilities, include pathological agents that are well recognized as having a negative effect on pregnancy or other pathogens for which the involvement in fertility problems is unclear. However, analyzing the literature, it appears that the most prevalent causes of infertility in pedigree cats may well be, in females, uterine pathology (sub-clinical endometritis, cystic endometrial hyperplasia, or mucometra), and in males, teratospermia, which may be "permanent" and linked to reduced heterozygosity or "transient" in tomcats that present normal quantitative spermatogenic parameters. The influence of the breed is unknown. There is an urgent need to study the origin of infertility in purebred cats in order to improve the diagnosis and to develop treatments that may restore normal fertility in subfertile or infertile animals.

Small animal reproduction: Scientific facts versus dogmas or unverified beliefs.

Small Animal Reproduction is a rather recent topic in the field of Animal Reproduction. Although some continuing educational societies aim to improve the level of general knowledge, published scientific facts are still limited, and there are many hypotheses or affirmations that remain unverified or even sometimes dogmatic or empirical. Through examples, this article reviews the main causes that may sometimes challenge a veterinary practitioner faced with a dog or a cat presenting reproductive problems. It could be difficult to orientate the diagnosis or the therapeutic approach, not knowing if what is found in the literature is a scientific proof or just an unverified affirmation. This problem may result from the fact that there are still physiological processes that remain unclear, but also that some common beliefs are based on old studies that have not been repeated or verifiedIn addition, to make it more challenging to seek for the information, there are unverifiedaffirmations,unclear nomenclature, clinical conditions which have not been standardized in the literature, clinical conditions mostly described through case reports, lack of studies on specific topics, areas of small animal reproduction that have been neglected by researchers, contradictory data or even studies that lack objectivity. The growing interest for research in Small Animal Reproduction will probably reduce the gap between unverified beliefs and scientific facts.

Contraceptive implants used by cat breeders in France: a study of 140 purebred cats.

OBJECTIVES: Deslorelin 4.7 mg and melatonin 18 mg subcutaneous implants were studied in purebred male and female cats, via questionnaires sent to French cat breeders, to assess breed, age, duration of the contraceptive effect, fertility after use, changes in behaviour and side effects. METHODS: Reproductive data were collected in 57 tom cats and 41 queens implanted with deslorelin 4.7 mg, and 42 queens implanted with melatonin 18 mg, for a total of 140 purebred cats, from 38 different catteries, representing 18 breeds. RESULTS: Using deslorelin (Suprelorin 4.7 mg; Virbac), sexual behaviour in males was inhibited for a mean ± SD of 13.4 ± 3.2 (range 8-21) months in 37/57 cats. Of these, 24/37 mated successfully and produced litters at a mean of 15.5 ± 3.6 (range 9-20) months. Inhibition lasted 11 ± 1.1 (range 9-12; n = 6), 13.2 ± 2.4 (range 12-18; n = 6) and 15 ± 3.5 (range 9-18; n = 6) months in Norwegian Forest Cat, Singapura and Ragdoll males, respectively. In 26/41 females implanted with deslorelin 4.7 mg, oestrus was inhibited for a mean of 16.0 ± 5.7 (8-38) months; 12/26 went on to produce a litter. Of the side effects specific to females: two presented persistent oestrus, leading to the removal of the implant; two developed lactation; one had fibroadenomatosis; and one was sterilised owing to cystic endometrial hyperplasia. Using melatonin (Melovine 18 mg; Ceva), 33/42 females had oestrus inhibited for a mean of 86 ± 50 (range 21-277) days after implantation with a peak return to oestrus in March, and 12/33 had a subsequent litter. No side effects were reported. CONCLUSIONS AND RELEVANCE: This study is the first to collect a large amount of field data, in 140 purebred male and female cats where a deslorelin 4.7 mg or a melatonin 18 mg implant was used. These field results may allow for more accurate clinical advice and open up new avenues of research.

The View of the French Dog Breeders in Relation to Female Reproduction, Maternal Care and Stress during the Peripartum Period.

In France, as in many other western countries, dogs are an important part of the society as pets or working animals. The exact demand for puppies in France is unknown, as is the proportion of dogs coming from different breeding sources. Nevertheless, the origin of puppies is important since young dogs from puppy mills and pet stores appear to be more likely to develop behavioral disorders. Puppies coming from responsible breeders, on the other hand, tend to be better adapted. In well-managed kennels, an explanation for these behavioral differences may be associated with greater contact of litters with the dam and humans. Another factor that might influence a dog's temperament and character is maternal behavior. Although recent studies have shown that the quality of maternal care in dogs is important, direct effects on the development of behavioral problems such as fearfulness or noise sensitivity are still controversial. To better understand the view of breeders, due to an increased interest in maternal care of dogs, an online questionnaire was developed to assess the dog breeders' profiles and to explore their perception of normal maternal and stress-related behaviors during the peripartum period. In addition, the management of the female during this critical time was queried. Three-hundred and forty-five French dog breeders, representing 91 breeds, completed the online survey. Considering the demographics of the responders, breeding activity in France is mostly family-based with 76% raising two breeds of dogs that produce, on average, five litters/year. Around 60% of the breeders use progesterone levels to determine breeding date. The whelping date is estimated using multiple criteria and most labors and deliveries happen under human supervision. The majority of behaviors associated to good maternal care are defined by the vast majority as being related to more attention of the bitch towards the puppies with the frequency of nursing and licking being important clues. Globally, the peripartum is perceived as a stressful period and to minimize stress and reassure the bitch the favored method used is increasing human presence. Problems related to maternal behavior were described, notably with primiparous bitches.

Influence of dog-appeasing pheromone on canine maternal behaviour during the peripartum and neonatal periods.

BACKGROUND: Parturition and the initial postpartum period are important moments in the reproductive cycle of dogs. METHODS: A study assessed the effect of ADAPTIL, a dog-appeasing pheromone, on maternal behaviour during peripartum. Bitches were continuously exposed to ADAPTIL (n=20) or placebo (n=21) in double-blinded conditions from an average of about seven days before parturition up to 21 days postpartum. Differences in maternal behaviour in relation to the treatment were evaluated by the observation of specific activities through video recordings, such as the time spent by the bitch in close contact with the puppies, oronasal interaction and nursing duration and position. Videos were recorded at four time points (W0: within the first 48 hours of whelping; W1: one week after parturition; W2: two weeks after parturition; and W3: three weeks after parturition). In addition, the perception of breeders in relation to the quality of maternal care, puppies' wellbeing and overall relationship between the bitches and the puppies was evaluated using Visual Analogue Scale at the same time points. Moreover, the daily activity of the bitches was measured by using an electronic device (FitBark dog activity trackers, Kansas City, Missouri). RESULTS: For all observed maternal behaviours, there was a steady decrease in levels as the puppies developed, independently of treatment. However, bitches exposed to ADAPTIL tended to nurse significantly more in lying position, while those exposed to the placebo nursed more in a seated position, especially at W1 (P=0.06) and W3 (P=0.005). According to the breeders, the attention scores of bitches towards puppies were significantly higher in ADAPTIL than in the placebo group at each time point (P=0.01). Moreover, a difference according to parity was observed (P=0.004), with greater attention score displayed by primiparous bitches exposed to ADAPTIL compared with placebo on W0 (P=0.02), W1 and W3 (P<0.001). The global mother-puppies relationship was also perceived as significantly better (P=0.0002) by breeders of bitches exposed to ADAPTIL, with significant differences at W2 (P=0.01) and W3 (P=0.001). The bitches' daily activity increased starting two days before the whelp, peaked during parturition and then gradually declined up until four days postpartum. There was a trend towards a difference in the activity level according to the treatment during the full study period (P=0.09) and at two days before parturition (P=0.07). Bitches exposed to ADAPTIL were more active compared with placebo in relation to the FitBark data. CONCLUSION: The use of ADAPTIL in maternity modulated maternal behaviours. Concerning the caregiver's view, bitches under the influence of ADAPTIL had greater and extended attention towards the puppies and they were eager to stay with the puppies for a longer time.

Neonatal puppy survival after planned caesarean section in the bitch using aglepristone as a primer: A retrospective study on 74 cases.

Since most of dystocia end up in caesarean sections (C-sections), the history of any problem during whelping is a good reason to plan in advance a further C-section. Our aim was to confirm that on a large sample and over an extended period of time, mortality in puppies <2 weeks of age was low, born after a planned C-section using aglepristone as a primer. Seventy-four C-sections on 59 different bitches were included. Bitches were monitored during oestrus to estimate the day of ovulation by progesterone assays; 60, 61 or 62 days after ovulation, foetal viability was checked by ultrasonography and progesterone plasma level was measured. None of the bitches was at term (progesterone plasma level >2 ng/ml). An injection of aglepristone was performed in late afternoon to block the effect of progesterone, mimicking its drop at the end of pregnancy. The C-section was conducted the following morning. Twenty-one breeds were represented most of which were bulldogs (26%, 21/74) and Great Danes (16%, 13/74). Four hundred and thirty-five puppies were born. A total of 43/435 puppies died within the first 2 weeks (9.89%). None of the puppies showed any external signs of prematurity. The average number of deaths per litter relative to the date after ovulation was similar (0.5 pups per litter at day 60, 0.7 at day 61, 0.4 at day 62). This study shows that planned C-section after an accurate determination of ovulation and using aglepristone as a primer is a safe procedure for bitches and their offspring. It may be offered to owners if a pregnant bitch is "at risk" of dystocia.

Manipulation of the oestrous cycle of the bitch-what works for now.

Manipulation of the oestrous cycle of the bitch is a common request of pet owners for different reasons. On one side, clients may be interested in temporally preventing oestrus until the bitch has finished the performance career, or to manage the kennel in the presence of a stud dog and to avoid undesirable breeding. Additionally, clients with no interest to use their animal for breeding, may, for ethical reasons, prefer to avoid elective surgery such as ovariectomy, to control reproduction. On the other side, clients may want to induce oestrus to better control the time of birth, to use a male with limited availability during a specific time, or to reduce the duration of the interoestrous interval in bitches with a long oestrous cycle (>8 months). Both these options for oestrous manipulation are based on hormonal treatments. For oestrous prevention, synthetic progesterone has been used since the 1960s. Induction of oestrus has been established with reliable results using dopamine agonists in the late 1980s and more recently using GnRH agonists. This review is focusing on the current and recently available treatments to manipulate the oestrous cycle in the bitch.

Is electroejaculation a safe procedure in cats? An endoscopic and histological prospective blinded study.

Transrectal electrostimulation is a technique used for semen collection in several animals including felids, but also in humans with spinal cord injuries and in specific cases of anejaculation. To the authors' knowledge, safety and innocuity on rectal mucosa has not yet been assessed histologically. A prospective study was performed on 10 European cats in order to determine the impact of electroejaculation on the rectal mucosa, by performing endoscopic and histological evaluation of the rectal mucosa. Endoscopic evaluation was performed before and after semen collection. Standardized biopsy specimens were collected immediately after semen collection, on the site of electrostimulation and on the proximal part of the descending colon as a control, for further double-blinded histopathological evaluation. The degree of inflammation was evaluated following the criteria from the WSAVA Gastrointestinal Standardization Group. Semen collection by electrostimulation did not induce significant histological and endoscopic lesions. Electrostimulation may therefore be considered as a safe technique to collect semen in cats.

Progesterone plays a critical role in canine oocyte maturation and fertilization.

Canine oocyte maturation and fertilization take place within the oviducts under increasing plasma levels of progesterone (P4). In order to investigate the role of P4 in these processes, 51 beagle bitches were treated with the P4 receptor antagonist aglepristone at the end of proestrus and 32 females were kept untreated. Fifteen treated and 13 control bitches were inseminated at Days +1 and +2 after ovulation (Day 0). Stages of oocyte maturation and embryo development were determined after ovariectomy at different time points after ovulation. Aglepristone did not prevent ovulation but delayed the resumption of oocyte meiosis and inhibited its progression: first metaphase I (MI) stage was observed at 173 h postovulation and 39% of oocytes reached MII as late as 335 h postovulation in treated females whereas first MI occurred at 76 h and 100% of oocytes were in MII at 109 h postovulation in controls. Aglepristone extended the stay of morphologically normal oocytes within the oviducts: first signs of oocyte degeneration were observed at 335 h in treated versus 100- to 110-h postovulation in control bitches. In inseminated females, aglepristone prevented sperm progression toward the oviducts and fertilization, although motile spermatozoa were observed in the uterine tip flush and within the cranial uterine glands. A proteomic analysis of the tubal fluid from treated and control noninseminated bitches at Day +4 found evidence of 79 differential proteins potentially involved in the oocyte phenotype. In conclusion, P4 plays key roles in postovulatory canine oocyte maturation, aging, and in fertilization.

Findings in uterine biopsies obtained by laparotomy from bitches with unexplained infertility or pregnancy loss: an observational study.

Surgical uterine biopsies (SUBs) were obtained by laparotomy from 14 bitches with unexplained infertility whose pregnancy was not confirmed (NCP group) and from 7 bitches that had experienced unexplained pregnancy loss (PL). SUBs were obtained during the luteal phase of the cycle in 16 of 21 bitches. Overall, the biopsied uterine tissues presented obvious alterations in 17 of 21 cases. In the NCP group, 11 of 14 bitches had varying degrees of uterine lesions. In this group, six of nine bitches with macroscopically normal uteri presented histopathologic changes in the endometrium. All bitches with macroscopic modifications (five of 14) presented endometrial changes. Cytologic examination of the uterine fluid revealed eosinophilic amorphous material, erythrocytes, and a low number of degenerated leukocytes. On histopathologic examination, the most common lesion was fibrosis with degeneration of the endometrial glands (FDEGs six of 11), followed by endometritis (four of 11), cystic endometrial hyperplasia (CEH) (two of 11) and pseudoplacentational endometrial hyperplasia (PEH) (two of 11). Other lesions were adenomyosis, mucometra, and an endometrial polyp (one each of 11). Mixed lesions were present in four of the 11 bitches. In the PL group, macroscopic modifications consisted of intraluminal uterine dilatation (seven of seven) and presence of ovarian cysts (three of 7). Cytologic examination of the uterine content revealed high leukocyte counts. Histopathologic endometrial changes were found in six of seven bitches and included endometritis (three of seven), PEH (two of 6), pyometra (one of six), and CEH together with mucometra (one of six). Research of infectious agents was performed in 20 of 21 cases. Surprisingly, no bacteria or viruses were isolated from the uterine lumen in any of the cases. When the SUB was performed in diestrus, a medical treatment with aglepristone or with prostaglandins was established to avoid the risk of postoperative-induced pyometra. One case of complete uterine stenosis was the only side effect observed in the bitches that were not ovariohysterectomized after the collection of a biopsy (17 of 21). In conclusion, the most common findings in our biopsy specimens were FDEGs, endometritis, and hyperplastic remodeling of the endometrium (CEH and PEH). SUBs might provide useful information regarding the cause of infertility or PL. This technique allows visual inspection and palpation of the whole uterus and ovaries before the selection of a precise site for the biopsy. The size of the tissue sample collected can be large enough for an accurate diagnosis and samples for research of infectious agents can be obtained with no risk of contamination. Risk of side effects seems low when SUBs are performed according to this protocol, including postsurgery treatment with aglepristone or prostaglandins.