Proliferation and apoptosis studies of interplacental areas after aglepristone treatment for planned cesarean section in pregnant bitches.

Background and Aim: Progesterone (P4) is the main hormone for pregnancy maintenance, occurring approximately 62-64 days after ovulation in bitches. Progesterone acts by binding to specific receptors. Aglepristone is a progesterone receptor (PR) antagonist with a higher affinity for PR binding. There are no published studies on cell proliferation and apoptosis in the canine uterus at the time of parturition. Therefore, this study aimed to determine the local effects of aglepristone on cell proliferation and apoptosis of interplacental uterine tissue during planned cesarean section (C-section) in bitches. Materials and Methods: In this study, 13 client-owned French bulldogs were examined. Bitches were divided into treatment (n = 8) and control (n = 5) groups. Ovulation timing was predicted based on the serum P4 level on 62-64 days post-ovulation for parturition. Serum P4 levels were measured before (on 60-day post-ovulation) and on C-section day (on 61-day post-ovulation). Aglepristone (Alizine®), 15 mg/kg subcutaneously (SC), was administered on 60 days post-ovulation in the treatment group. A C-section was planned 20-24 h later, and interplacental uterine areas were collected from both groups during the C-section. Immunohistochemistry based on Ki-67 and TUNEL assay was used to evaluate cell proliferation and apoptosis in four different interplacental uterine tissue layers (epithelium, stroma, glandular epithelium, and myometrium). Data are reported as mean ± standard deviation. Kruskal-Wallis test was used for comparisons of more than two independent groups. P value of 0.05 was considered statistically significant. Results: One bitch in the treatment group was excluded due to emergency C-section 8 h after aglepristone administration. Serum P4 levels (ng/mL) at 20-24 h before and at C-section were 6.09 ± 2.72 and 4.32 ± 2.2 in the treatment group (n = 7) and 5.45 ± 1.28 and 3.67 ± 1.89 in the control group (n = 5), respectively. Proliferation (PI) and apoptotic (AI) indices were <5% and >45%, respectively, in both the treatment (n = 5) and control (n = 3) groups. PI and AI were detected at interplacental areas. Conclusion: There were no significant differences in serum P4 levels or PI and AI indices between the groups. The PI <5% and AI was higher than 45% in both groups. Aglepristone did not have a direct effect on the serum P4 levels in both groups. These results correlated with the natural physiology of parturition preparation. Aglepristone 15 mg/kg SC injected 20-24 h before parturition had no effect on the P4 level, nor were any harmful effects observed for a planned C-section in pregnant bitches.

Aerobic Vaginal Microflora in Gestational and Non-Gestational Bitches (Canis lupus familiaris).

The vaginal tract comprises commensal microorganisms, which play an essential role in the health of the reproductive tract. Any dysbiosis in the vaginal microenvironment may lead to severe urinary tract infections or even infertility. This study aimed to evaluate the aerobic bacterial flora isolated from vaginal samples from 100 lactating bitches in the antepartum period (n = 3), postpartum period (n = 80), and with Lactatio sine graviditate (n = 17). Before vaginal swabs, all the bitches went through a gynecology consult, along with milk and blood sampling. Standard microbiological techniques were used for bacterial isolation. Among the 100 vaginal samples analyzed, 82% had a positive microbiological outcome, while 18% were negative. The microbiologic profile listed 17 different genera. The main isolated bacterial families were Micrococcaceae, Staphylococcaceae, Morganellaceae, Bacillaceae, and Rhizobiaceae. At the same time, strains like Agrobacterium radiobacter, Ochrobactrum anthropi, Chromobacterium violaceum, Burkholderia mallei, Bacillus pumllus, or Staphylococcus xylosus were isolated for the first time from the vaginal secretion of lactating bitches. The microbiological data demonstrates that lactating bitches' vaginal discharge is heterogeneous and may be affected by coitus, sampling season, age, and reproductive status.

Clinical, Ovulatory and Histological Effect of a Postnatal Testosterone Treatment in Female Dogs.

It has been described in some female mammalian species that postnatal androgenization causes reproductive structural and functional abnormalities. The objective of this study was to describe the clinical, ovulatory and genital effects of postnatal androgens in female dogs. Ten newborn female crossbred puppies were randomly assigned to: testosterone enanthate 18 mg/100 g sc (TE; n = 5) or placebo sc (PL; n = 5). The puppies were physically followed up until puberty when ovulation was tested by serum progesterone. Then, ovariohysterectomies were performed, and genital tracts were grossly and histomorphometrically examined. At puberty, all the females had normal estrous behavior and ovulated without age and body weight differences. All TE puppies presented mild clitoris enlargement. Gross and microscopical ovarian examination did not reveal differences. Conversely, the endometrial area occupied by uterine glands as well as the height of the glandular and luminal epithelium were higher in the TE than in the PL group (<0.01). The height of the endometrium and myometrial thickness did not differ between groups. It was concluded that a supraphysiological postnatal dose of testosterone did not affect ovulatory capacity, nor did it provoke gonadal histological alterations, although it caused an increased area of endometrial glands and a higher uterine epithelium.

The Canine Vaginal Flora: A Large-Cohort Retrospective Study.

Microbiological examinations are frequently performed as part of breeding management examinations in the bitch, but also in case of (suspected) reproductive tract problems. As most bacteria are opportunistic pathogens, evaluation of bacterial findings is challenging for veterinarians. Besides, breeders might request antimicrobial treatment in breeding bitches, fearing conception failure-even without medical indication. Considering the rising threat of antimicrobial resistance, gaining deeper insights into the bacterial findings from the vagina of healthy and (suspected) reproductive-diseased bitches might contribute to the knowledge of the canine aerobic vaginal flora and consequently improve the responsible use of antibiotics. We analyzed results from bacteriological cultures of 23,254 vaginal swabs sent in to three commercial laboratories in Germany between 2015 and 2021, where standard aerobic microbiological examination was carried out. We found a variety of 319 bacterial species that mostly grew in mixed cultures of two or more bacterial species. Commonly found species were Escherichia coli, beta-hemolytic Streptococci, coagulase-positive Staphylococci, Pasteurellales, and aerobic sporulators, as well as other Streptococcus spp. Our results showed a large diversity of the canine vaginal flora in healthy and (suspected) reproductive-diseased bitches. They largely support earlier findings of small studies on the physiological canine vaginal flora, emphasizing that solely the results of a bacterial evaluation should not be the base for antimicrobial treatment. Instead, bacterial findings should be evaluated with the results of a clinical gynecological examination.

Relationships between haptoglobin and inter-alpha trypsin inhibitor heavy chain 4 levels in local and peripheral blood and systemic inflammatory response syndrome in bitches with pyometra.

We aimed to assess the usefulness of monitoring inter-alpha trypsin inhibitor heavy chain 4 (ITIH4) and haptoglobin (Hp) in peripheral and local blood in canine pyometra, and evaluation the relationships among acute phase proteins (APPs), systemic inflammatory response syndrome (SIRS) and the presence of bacteria. The material was collected from bitches with pyometra and from healthy ones. Blood was taken from the cephalic and uterine veins. APPs levels were quantified by ELISA. In the peripheral circulation, the Hp was higher in animals with open-cervix pyometra (OCP) than in the closed-cervix pyometra (CCP) and the control group. The Hp concentration was not correlated with age, with the presence of SIRS or with the type of bacteria (Gram-negative, Gram-positive or mixed flora). The ITIH4 concentrations in the peripheral blood did not differ significantly in the cases of pyometra. The Hp concentration in the local circulation increased in the OCP but not in the CCP groups, although the histopathological changes in the endometrium were similar. Peripheral Hp concentrations may be a useful tool in differentiating between the types of pyometra.

Vaginal aerobic bacteria of healthy bitches and those with fertility problems.

The most common problems in veterinary practice in bitches are bacterial infections of the reproductive tract associated with fertility problems. Research to determine the correlation between the health status of female dogs and bacterial flora of the genital tract has been ongoing for years, but the results obtained by different authors are often contradictory, and do not always concern breeding bitches. Our study identified the most common aerobic bacteria in the genital tract of numerous breeding bitches population. A total of 275 breeding dogs in anestrous phase of the estrous cycle were included in this study. 198 were qualified to the first group with no genital tract infections and no reproductive disorders. 68 bitches were qualified to the second group with complications such as: infertility, abortion, foetus resorptions and newborn mortality. The type of bacterial isolates was almost the same in the healthy bitches and the group with fertility problems. The most common bacteria obtained from the vaginal tract of the tested dogs were Streptococcus spp., Staphylococcus spp., Mycoplasma canis and Escherichia coli. There were no significant differences in bacterial prevalence in the group with reproductive problems versus healthy dogs; however, we found a statistically significant difference between both groups when the numbers of bacterial strains were compared. The number of one-strain bitches was statistically higher in the problematic group than in the non-problematic one. Bacterial culturing of vaginal swab specimens from breeding bitches without clinical signs of genital disease is of little value. Furthermore, it should always be preceded by an examination (clinical, cytological or vaginoscopy etc.). The request or requirement to perform vaginal cultures that is made by some breeders, while common, is not diagnostic for any pathologic condition and the results of these cultures should never be used to determine if antibiotic therapy is indicated.

Microbiological and hematological aspects of canine pyometra and associated risk factors.

Canine Pyometra, also known as cystic endometrial hyperplasia complex, is a common reproductive issue in bitches. This study aimed to identify associated risk factors, hematological variation, bacteria involved, and the most potent anti-bacterial against bacterial isolates of canine pyometra. Forty-five bitches of different habitats, breeds, and ages infected with pyometra were included in the study. The samples were cultured to isolate bacteria associated with the pyometra and antibiotic sensitivity was done for each bacterial isolates to get antibiogram. The study findings showed that potential risk factors such as age group, medroxyprogesterone acetate administration, and changes in the white blood cells parameters were significantly associated (P < 0.05) with the type of pyometra. Closed cervix pyometra in dogs showed significantly higher prevalence of clinical signs including depression, vomiting, abdominal enlargement, and fever compared to the open cervix pyometra. Low levels of red blood cells, pack cell volume, and hemoglobin indicated that the pyometra-infected dogs were more likely to have normocytic, normochromic, and non-regenerative anemia. Pyometra was attributed to an increase in AST (Aspertate aminotransferase), ALT (Alanine transaminase), ALP (Alkaline phosphatase), BUN (Blood Urea Nitrogen), and Creatinine while a decrease in serum albumin. Of the all bacterial isolates, E. coli (35.55%) was the most common pathogen isolated from canine pyometra, followed by Pseudomonas spp. (26.66%). E coli and Pseudomonas spp. were susceptible to Imipenem, Amikacin, and Gentamicin while highly resistant to Ampicillin and Erythromycin. Imipenem, Amikacin, and Gentamicin were the most sensitive antibiotics, while Ampicillin and Erythromycin were the most resistant antibiotics for the bacterial strain isolated from canine pyometra. Multidrug resistant was observed in 26 of the isolated bacteria, indicating acquired resistance due to improper and uncontrolled use. Hence early diagnosis and close monitoring of antimicrobial susceptibility before therapeutic intervention is indispensable in preventing the global threat of antimicrobial resistance.

Uterine dynamics, blood profiles, and electronic fetal monitoring of primiparous and multiparous bitches classified according to their weight.

Perinatal mortality occurs in all species. In dogs, mortality rates have been reported to range from 5 to 35%. Electronic fetal and uterine monitoring has recently been used in domestic animals to monitor the mother and newborn before and during parturition. In this way, the fetal heart rate and uterine dynamics can be monitored. This study evaluated the uterine dynamics of bitches with different weights and parity. Ninety-six bitches and their 476 puppies were divided into four experimental groups containing 24 individuals each (12 primiparous bitches and 12 multiparous bitches), according to body weight: G1 (4-8 kg), G2 (8.1-16 kg), G3 (16.1 to 32 kg), and G4 (32.1 to 39.6 kg). The fetal heart rate decelerations (dip 2 patterns), uterine dynamics, and bitches' blood profiles were evaluated, including levels of glucose, lactate, pCO2, pO2, pH, HCO3-, and Ca++. The dam weight can affect the vitality of newborns and the uterine dynamics, with differences in the frequency, intensity, and duration of myometrial contractions. The expulsion interval between puppies was longest in primiparous bitches with low weight and shortest in multiparous bitches with high weight. The expulsion interval and the number of stillborn females were higher in primiparous bitches with high weight. Newborn male puppies were significantly heavier than newborn females.

Uterine prolapse in a non-pregnant bitch.

A ten-year-old mixed breed bitch was presented for a tissue prolapse protruding from her vulva. Following detailed examination and stabilization, the ovaries and uterine horns were removed by laparotomy, whereas the prolapsed tissue identified as uterus including cervix was removed vaginally. Histology confirmed uterine prolapse, a rare condition in bitches usually found shortly after birth especially due to dystocia. In contrast, the present case was found in a nulliparous non-pregnant bitch. Diagnostic and therapeutic approaches, including microbiological and histological findings, are described and discussed critically.

A rare case of colonic diverticulum in a Bitch.

Abstract. Background: Colonic diverticulum is one of the rare findings in dogs characterized by an out-pouching of mucosal and submucosal layers through the defect in muscularis layer of the colon. Case description: A five years old intact female Labrador was presented with an anamnesis of dyschezia and tenesmus. Findings/treatment and outcome: Rectal examination was normal, and the survey radiograph showed an almost crescent shaped abnormal dilatation (10.52 cm × 6.21 cm) with gas and increased radiopaque material, dorsal to the urinary bladder and ventral to the descending colon suggesting fecal stasis. Ultrasonographic examination revealed gas-filled out-pouching with hyperechoic colon wall and acoustic shadowing. Exploratory celiotomy confirmed the diagnosis of colonic diverticulum, and diverticulectomy was performed. All four layers of the colonic wall were detected histopathologically in the biopsy sample and excluded neoplasia. The dog recovered uneventfully with no post-operative complications. Conclusion: This surgery produced an excellent resolution of clinical signs. To our knowledge, this is one of the few cases of colonic diverticulum reported in dogs.

Clinical retrospective study of vaginal hyperplasia in the bitch (2012-2022).

A retrospective study was used to analyse canine vaginal hyperplasia ambulatory cases in Small animal clinic of the University Veterinary Hospital of the Faculty of Veterinary Medicine, Trakia University in Stara Zagora, Bulgaria between 2012 and 2022. The study was done to investigate the morbidity of vaginal hyperplasia in the bitch and its relationship with the breed, body weight, age, stage of the oestrous cycle, parity of dogs and grade of the condition. Grade II and grade III vaginal hyperplasia were observed in 3.11% of all female dog genital pathological cases. Twenty-eight canine breeds were affected and the largest number were Central Asian Shepherds (30.47%). Most frequently the dogs were between 2 and 3 years old (29.68%), above 40 kg body weight (58.59%) and during follicular phase (proestrus and oestrus) of the oestrous cycle (82.02%). All the affected bitches were non-spayed (100%) and nulliparous were 70.31% of them. Grade III vaginal hyperplasia was more frequently observed (53.13%) than grade II of the condition. High correlation was found between age of the bitches and grade of the vaginal hyperplasia (R2 = .551; SEE = 0.053), body weight and grade of the condition (R2 = .820; SEE = 0.054), and age of the animals and their body weight (R2 = .820; SEE = 1.27).

Clinical and ultrasonographic findings of ovarian tumours in bitches: A retrospective study.

A retrospective study was carried out to investigate incidence, clinical signs and ultrasonographic findings of ovarian tumours in a population of dogs referred to the Veterinary Teaching Hospital of the University of Perugia (Italy) and Anicura Tyrus Veterinary Clinic (Terni, Italy). The period of study ranged from January 2005 to December 2021. A total of 1910 dogs were affected by neoplasia but only 35 of them (1.8%), of different breeds and ages, were found to have ovarian tumours. Ultrasound of the ovaries was performed based on clinical signs; the diagnosis was achieved after ultrasound findings prompted ovariohysterectomy and ovarian pathologic evaluation In our study, the age of bitches affected by ovarian neoplasia ranged from 3 to 20 years (mean 9.6 ± 3.8). The histopathological findings of ovarian masses identified 16 granulosa cell tumours (GCT) (46%), 7 adenomas (20%), 5 adenocarcinomas (14%), 2 teratomas (6%), 1 leiomyoma (3%), 1 luteoma (3%), 1 tecoma (3%), 1 dysgerminoma (3%), and 1 haemangiosarcoma (3%). In particular, with respect to clinical signs, 69% of bitches showed abnormalities of estrus cycle (short interestral interval, persistent estrus, prolonged interestral interval). The other main clinical signs included abdominal distention, palpable abdominal mass, vulvovaginal discharge, polyuria/polydipsia, mammary masses. When present, the laboratory abnormalities were slight anemia and leucocytosis with neutrophilia. The tumours were ultrasonographically classified as mainly solid: 12/35 (34%) (1 adenoma, 4 adenocarcinomas, 1 dysgerminoma, 1 haemangiosarcoma, 1 leyomioma, 1 luteoma, 1 GCT, 1 tecoma, 1 teratoma); solid with cystic component 13/35 (37%) (9 GCT, 2 Adenomas, 1 adenocarcinoma, 1 teratoma); and mainly cystic 10/35 (29%) (6 GCTs, 4 adenomas). In our study, the ultrasound examination allowed us to suspect ovarian neoplasia in asymptomatic subjects referred for breeding management or for preventive health check. On the basis of our data, we proposed to perform a complete periodic examination of the reproductive system once a year from 6 years. Nevertheless, the presence of ovarian neoplasms found in young subjects, during breeding management, suggest including routine ultrasound examination of the reproductive tract.

Factors Affecting Pregnancy Success in the Bitch Following Transcervical Insemination.

Endoscopic-assisted transcervical inseminations (TCIs) have become increasingly popular. The aim of this retrospective clinical study was to evaluate data from the TCIs performed at our facility. We evaluated data from January 2018 through December 2021. This included 137 cases with fresh, 67 cases chilled, and 63 cases using frozen-thawed semen. All bitches underwent breeding management to determine the ideal breeding period. All semen samples were evaluated for total number of sperm, total motility, and progressive motility. Pregnancy was determined by B-mode ultrasonography about 4 weeks after the breeding. Litter size was determined by radiographs performed around the last week of gestation. The pregnancy rate was 83.21% for fresh, 67.16% for chilled, and 66.67% for frozen-thawed semen. There was a significant difference in litter size between fresh semen (6.82 puppies per litter) and both chilled (5.21 puppies per litter) and frozen-thawed (4.59 puppies per litter) semen (P < .05). There was no significant difference in litter size between chilled and frozen-thawed semen. There was no difference in pregnancy rates between clinicians performing the inseminations. Pregnancy rate was not different when sedation was used for the insemination (66.67%) compared to when sedation was not used (74.84%; P > .05). Performing 2 TCIs during the fertile period, regardless of the semen type, resulted in an increase of 6.6% in pregnancy rate (P > .05) and an increase of 0.7 puppies per litter, on average (P > .05). These results can be used to help guide recommendations for breeding clients on the best options to increase both pregnancy rate and litter size for their breeding.

A Prospective Cohort Study Investigating the Impact of Neutering Bitches Prepubertally or Post-Pubertally on Physical Development.

No previous large prospective cohort studies have been identified that have investigated the impact of the surgical neutering of bitches before or after known puberty on their growth and physical development. This study was designed to examine the data on physical development, vulval size, and conformation for bitches neutered by ovariohysterectomy before puberty (PPN, n = 155) or after puberty (control, n = 151) using a prospective cohort study design. Data were gathered at six- and 17-months of age using bespoke physical assessment forms and digital images of the vulva. PPN bitches had greater changes in height measurements (mean difference = 2.039, SEM = 0.334, 91% CI = 1.471 to 2.608, p < 0.001) and smaller changes in the measurements of vulval length (mean difference = -0.377, SEM = 0.079, 91% CI = -0.511 to -0.243, p < 0.001) and width (mean difference = -0.221, SEM = 0.063, 91% CI = -0.328 to -0.113, p < 0.001) between six- and 17-months of age than for the control bitches. Although not significant, the PPN bitches were taller (mean 58.5 vs. 56.6 cm) and heavier (mean 28.3 vs. 27.3 kg) with smaller vulval size measurements (mean vulval length 2.8 vs. 3.2 cm, mean vulval width 1.7 vs. 2.1 cm) at 17-months of age. At 17-months of age, significantly more PPN bitches had vulvas that appeared juvenile (Yates' Chi-square = 14.834, D.F. = 1, p < 0.001) and recessed (Yates' Chi-square = 7.792, D.F. = 1, p = 0.005) at the physical assessment, and significantly more PPN bitches had vulvas that appeared 'recessed/inverted' on the examination of digital images (Chi-square = 9.902, D.F. = 1, p = 0.002). The results from this study suggest no contraindications to prepubertal ovariohysterectomy for large breed bitches. However, any longer-term health implications of these differences in physical development need to be investigated and better understood prior to recommendations being made.

Vincristine-associated total antioxidant and oxidant status of ovaries and in vitro nuclear oocyte maturation in dogs with canine transmissible venereal tumor.

The aim of this study is to evaluate the effects of scheduled vincristine sulfate therapy on canine oocyte quality and nuclear oocyte maturation, associated with total antioxidant and oxidant status of ovaries and Anti-Müllerian Hormone (AMH) concentrations in dogs with Canine Transmissible Venereal Tumor (CTVT). Six bitches suffering from CTVT and six healthy bitches were included in the study. Hemogram was carried out weekly. AMH measurements and ovariohysterectomy operations were performed after the termination of vincristine sulfate therapies. Tissue samples from ovaries were utilized for Malondialdehyde (MDA), reduced Glutathione (GSH), Superoxide Dismutase (SOD), Total Anti-oxidative Status (TAS), Total Oxidative Status (TOS) measurements, and Oxidative Stress Index (OSI) was calculated. Collected oocytes were evaluated for meiotic competence, after In Vitro Maturation (IVM) and parthenogenetic activation. No difference between the two groups was observed in hematologic parameters (P > 0.05). Meiotic stages of Germinal Vesicle Break Down (GVBD), Metaphase I (MI), and Metaphase II (MII) were significantly different between groups (P < 0.05). The number of oocytes reaching MII and meiotic resumption was lower in the CTVT group. Furthermore, AMH concentrations, oxidant parameters (OSI, TOS, and MDA), and antioxidant parameters (GSH, SOD, and TAS) were also statistically different between groups (P < 0.05). The results of this study show that vincristine sulfate application in the treatment of CTVT could alter oxidant/antioxidant status in ovaries. Apart from these, oocyte quality and IVM rates seem to decline related to gonadotoxicity. Moreover, AMH could be an important marker in the evaluation of oocyte qualities in bitches, as it is in women.

The effect of doxapram on survival and APGAR score in newborn puppies delivered by elective caesarean: A randomized controlled trial.

Doxapram is marketed as a respiratory stimulant and is used by some veterinarians to help with neonatal apnoea, especially in puppies delivered by caesarean. There is a lack of consensus as to whether the drug is effective and data on its safety are limited. Doxapram was compared to placebo (saline) in newborn puppies in a randomized, double-blinded clinical trial using two outcome measures: 7-day mortality rate and repeated APGAR score measurements. Higher APGAR scores have been positively correlated with survival and other health outcomes in newborns. Puppies were delivered by caesarean and a baseline APGAR score was measured. This was immediately followed by a randomly allocated intralingual injection of either doxapram or isotonic saline (of the same volume). Injection volumes were determined by the weight of the puppy and each injection was administered within a minute of birth. The mean dose of doxapram administered was 10.65 mg/kg. APGAR scores were measured again at 2, 5, 10 and 20 min. One hundred and seventy-one puppies from 45 elective caesareans were recruited into this study. Five out of 85 puppies died after receiving saline and 7 out of 86 died after receiving doxapram. Adjusting for the baseline APGAR score, the age of the mother and whether the puppy was a brachycephalic breed, there was insufficient evidence to conclude a difference in the odds of 7-day survival for puppies that received doxapram compared to those that received saline (p = .634). Adjusting for the baseline APGAR score, the weight of the mother, the litter size, the mother's parity number, the weight of the puppy and whether the puppy was a brachycephalic breed, there was insufficient evidence to conclude a difference in the probability of a puppy having an APGAR score of ten (the maximum APGAR score) between those that received doxapram compared to those that received saline (p = .631). Being a brachycephalic breed was not associated with an increased odds of 7-day mortality (p = .156) but the effect of the baseline APGAR score on the probability of having an APGAR score of ten was higher for brachycephalic than non-brachycephalic breeds (p = .01). There was insufficient evidence that intralingual doxapram provided an advantage (or disadvantage) compared to intralingual saline when used routinely in puppies delivered by elective caesarean and that were not apnoeic.

Isolation of extracellular vesicles from bitch's amnion-derived cells culture and their CD59 expression: Preliminary results.

Extracellular vesicles (EVs) are small spherical particles surrounded by a membrane with an unusual lipid composition and a striking cholesterol/phospholipidic ratio. About 2000 lipid and 3500 protein species were identified in EVs secreted by different cell sources. EVs mediate cell to cell communication in proximity to or distant from the cell of origin. In particular, it was suggested that they represent modulators of multiple processes during pregnancy. The aim of this study was to identify the presence of EVs in canine amnion-derived cells (ASCs) culture and the expression of CD 59 on their surface. Amniotic membrane was collected in PBS with antibiotics added from 2 bitches during elective caesarean section. Cells culture was prepared and EVs were isolated. EVs were used to evaluate CD59 expression by flow cytofluorimetry. We found that the majority of EVs expressed CD59. Our results could increase the knowledge about the complex mechanisms that regulate the pregnancy in the bitch.

Monitoring of canine pregnancy by considering Anti-Mullerian hormone, C-reactive protein, progesterone and complete blood count in pregnant and non-pregnant dogs.

In this study, we measured the serum concentration of anti-Mullerian hormone (AMH), C-reactive protein (CRP), progesterone (P4), and the complete blood count (CBC) in pregnant and non-pregnant bitches. The aim was to investigate the suitability of these parameters for monitoring canine pregnancy. Blood samples were collected from all bitches introduced for timed mating on the day of first mating (>5 ng/ml). The first blood sample after mating was obtained on day 12 post-copulation. The dogs whose pregnancy was confirmed on days 25 and 35, were allotted to the pregnancy positive group (G+) and those that were not pregnant were grouped as pregnancy negative (G). Ultrasonography (US) was performed on days 25, 35, 45 and 55 in pregnant (N = 13) and non-pregnant (N = 7) animals; The sonographic examinations in non-pregnant bitches were continued up to day 63, and in pregnant bitches they were also carried out one day after parturition (D+1). Blood samples were taken in parallel with these periods. Furthermore, the pregnant bitches were classified as G1A (1-2 puppies), G1B (3-4 puppies), and G1C (5-11 puppies) based on the number of puppies, and G1X (10 kg), G1Y (10-20 kg), and G1Z (>20 kg) based on their body weight. No significant difference was found between G+ and G-with regard to AMH, except on day 45, when AMH was higher in G+ (P < 0.01). On the other hand, the CRP values in the G+ exceeded those in the G-group on day 25 (38.26 vs. 15.66 mg/L, P < 0.05), on day 35 (32.54 vs. 15.97 mg/L, P < 0.05) and on day one after parturition (36.24 vs. 10.10 mg/L, P < 0.01). When puppy number was considered, it was discovered that CRP values significantly increased with puppy number on days 12 and 45 (G1A vs. G1B day 12: 4.13 vs 15.84 mg/L, P < 0.05; day 45: 12.40 vs. 25.76 mg/L, P < 0.001), and on day 35 (G1B vs. G1C: 24.18 vs. 38.87 mg/L, P < 0.01). With regards to AMH, this was only detectable on day 12 (G1A vs. G1B: 0.56 vs. 1.13 ng/mL, P < 0.05). When the body weight of the pregnant bitches was considered, bitches <10 kg had significantly higher AMH values than bitches bitch >20 kg on days 12 and 25 (day 12: 1.20 vs. 0.21 ng/mL, P < 0.01; day 25: 0.91 vs. 0.21 ng/mL, P < 0.05). This was not found in the case of CRP. The white blood cells (WBC) and the granulocytes (GRAN) were found to be higher in the G+ group (P < 0.01) on day 55, while the hematocrit (HCT) was significantly lower on day 45 (P < 0.05) and day 55 (P < 0.01). The increased GRAN was still detectable one day after parturition (P < 0.05). In conclusion, measurement of the AMH and CRP concentrations may contribute to determination of gestation stage and monitoring of the course of pregnancy; values are related to maternal body weight and number of puppies; however, AMH did not change over the course of a normal pregnancy. Sonography, the increase in CRP and complete blood count values may be beneficial for monitoring canine pregnancy. More studies are necessary to prove these findings.

A prospective cohort study investigating the peri- and postoperative outcomes following ovariohysterectomy in bitches neutered prepubertally or post-pubertally.

The aim of this prospective cohort study was to investigate perioperative and postoperative outcomes for bitches that were neutered at different times in relation to puberty (rather than age). Three hundred and six Labrador and Golden Retriever crossbreed bitches were neutered before (prepubertal neuter [PPN], n = 155) or after puberty (Control, n = 151) by experienced veterinarians at one of four veterinary practices (VP1-4). Data were gathered from veterinarians at the time of neutering and from bitch caregivers for the 14-day period after neutering using purpose-designed questionnaires. PPN bitches had shorter surgery durations than Control bitches (OR = 0.877, 91% CI = 0.800 to 0.961, P = 0.015). Longer surgery durations were reported for bitches that had intraoperative complications (OR = 1.620, 91% CI = 1.397 to 1.879, P < 0.001). Shorter surgery durations were reported for VP1 compared to all other veterinary practices (OR = 1.189 to 1.534, P < 0.001). There was no difference in the number of PPN or Control bitches that had perioperative complications (PPN: 3.9%, n = 6; Control: 7.7%, n = 11) and no bitch had more than one perioperative complication reported. For the 14-day postoperative period, fewer PPN (1.3%, n = 2) compared to Control bitches (8.9%, n = 13) had wound discharge (Chi-square = 11.151, D.F = 1, P = 0.001) or showed inappetence (PPN: 2.9%, n = 4; Control: 17.3%, n = 19, Chi-square = 14.884, D.F. = 1, P < 0.001). There was no difference in the number of PPN and Control bitches that had redness or swelling of the wound (52 PPN, 48 Control), showed attention to the wound (6 PPN, 6 Control), required veterinary attention (25 PPN, 23 Control), or that showed any other unusual behaviours: discomfort (27 PPN, 16 Control), uncharacteristic irritability (2 PPN, 4 Control), or uncharacteristic licking or chewing of parts of the body (15 PPN, 6 Control). Bitches neutered at VP2-4 were more likely to have wound redness or swelling reported than those neutered at VP1 (OR = 3.109 to 4.076, P = 0.003). Future studies should consider the impact of veterinary practice on results. These results for peri and postoperative complications could suggest that prepubertal ovariohysterectomy may be a suitable option for neutering bitches. However, it is recommended that they be interpreted together with evidence relating to the impacts on other factors including behaviour and longer-term health in order to make fully informed decisions.

Simultaneously administration of cabergoline and PMSG reduces the duration of estrus induction in anestrous bitches.

This study designed a protocol that would combine pregnant mare serum gonadotrophin (PMSG) and cabergoline (CAB) to induce estrus in bitches. Twenty clinically healthy adult and anestrous female dogs were randomly assigned into four groups. The first group was treated with 5.00 µg kg-1 CAB until the onset of proestrus or for 25 days. The second group was treated with 20.00 IU kg-1 PMSG for 5 days and 500 IU human chorionic gonadotropin (hCG) on the 5th day. The third group was treated with 5.00 µg kg-1 CAB for 10 days in combination with 20.00 IU kg-1 PMSG for 5 days and 500 IU hCG on the 10th day. The control group received 1.00 mL of normal saline. Ovarian changes were evaluated ultrasonographically, and the estrus cycle phase was examined by vaginal cytology. Respectively, three, three and four bitches showed clinical signs of proestrus in each treatment group. The intervals between treatment and proestrus for each group were 30.00 ± 3.05, 7.67 ± 1.20 and 13.00 ± 1.20 days, respectively. Two weeks after estrus, the progesterone mean was 14.51 ± 6.24, 19.96 ± 17.16 and 19.12 ± 9.26 ng mL-1 for each group, respectively. In ultrasonography examination, the largest follicle was identified at 15.66 ± 1.33, 11.66 ± 2.40 and 8.75 ± 2.17 days after the onset of proestrus and the largest follicle's size was measured 6.50 ± 0.55, 4.83 ± 1.64 and 7.07 ± 1.49 mm for each group, respectively. Although the combined use of CAB and PMSG reduced the duration of treatment, alteration of the duration or PMSG dosage can be helpful to improve the results.