Clinical Trial on the Usefulness of On-Site Evaluation of Canine Fetal Fluids by Reagent Test Strip in Puppies at Elective Caesarean Section

The reagent urinary test strips (TS) marketed for urines represent the first-line diagnostic tool in many instances. Therefore, the aim of the study was to assess the usefulness of TS for the on-field evaluation of fetal fluids’ composition in newborn dogs at elective caesarean section. Of a total of 137 puppies born at term, 127 survived and 10 did not survive. One hundred and thirteen amniotic and 107 allantoic samples from surviving newborns were collected, and 8 amniotic and allantoic fluids were collected from the non-surviving newborns and assessed by strips. Significantly lower amounts of amniotic glucose and higher amounts of amniotic and allantoic nitrites, amniotic protein, allantoic urobilinogen, and amniotic bilirubin concentrations were found in non-surviving when compared to surviving newborns. In the surviving ones, higher specific gravity and bilirubin concentrations, and lower pH, were found in allantoic than in amniotic fluids. Higher amniotic and allantoic glucose concentrations, higher amniotic and allantoic pH, and lower amniotic and allantoic protein concentrations were found in medium/large- than in small-sized puppies. The TS allowed the quick evaluation of fetal fluids in puppies at birth. The differences between surviving and non-surviving puppies seem to suggest that the on-site analysis of fetal fluids’ composition by TS could represent a first-line diagnostic tool in the field of canine neonatology, allowing the quick recognition of puppies needing assistance as a complementary tool for clinical evaluation.

Placenta Morphological and Amniotic Fluid Biochemical Changes Associated with Mid-Gestation Single and Twin Pregnancies in Red Sokoto Goats

Fetal fluids and placentas, which are important in feto-maternal homeostasis, could be affected by the number of developing fetuses. Therefore, placenta morphological and amniotic fluid biochemical changes associated with single and twin pregnancies in Red Sokoto goats were studied. Using Richardson’s formula, 2.1 × [CRL (cm) + 17], 12 intact gravid uteri (7 single-pregnant and 5 twin-pregnant) from mid-gestation [≈70-100 days gestational age (dGA)] pregnant goats were purposively selected. Feto-maternal biometrics [gravid uterine weight (GUW), total placental fluid volume (TPFV), average placentome diameter (APD), mean crown-rump length (MCRL), mean gestational age (MGA), mean fetal weight (MFW), total placentome number (TPN), and average interplacentomal distance (AID)]; amniotic fluid electrolytes [sodium (Na), potassium (K), chloride (Cl), calcium (Ca) and phosphorus (P)]; total protein; glucose; liver markers (AST and ALT); kidney markers (urea and creatinine), and cortisol concentrations were measured. The histology of the placentomes and the interplacentomal areas was also studied. The twin-pregnant (TP) goats had significantly higher (P<0.05) GUW, TPFV, APD, amniotic fluid glucose and cortisol; and lower (P<0.05) Ca levels. The rest of the parameters assessed did not differ between the TP and single-pregnant (SP) groups. Placentomes and the interplacentomal areas from the TP group had more diffuse villous inter-digitations and thicker luminal endothelium, respectively. It was concluded that TP Red Sokoto goats regulated placentome morphology, calcium, glucose and cortisol levels in order to achieve optimal conditions for the dams and the fetuses.

Integrins and OPN localize to adhesion complexes during placentation in sheep

Integrins and OPN are potential mediators of blastocyst attachment to the endometrium to initiate implantation. The goals were to examine the temporal/spatial pattern of expression of integrins at the endometrial–placental interface of sheep encompassing Days 9 through 80 of gestation and determine if OPN co-localizes with integrins. Results show the following: (1) αv, α4, β1, β3 and β5 integrins at the apical surface of endometrial luminal epithelium (LE) from Days 11 through 16 of pregnancy that indicate a role for these integrins during implantation; (2) large, intermittent aggregates of αv, α4, α5, β1 and β5 integrins at the endometrial–placental interface from Days 20 through 55, suggesting adaptation to a localized tissue remodeling stage of placentation; and (3) integrin adhesion complexes (IACs) containing αv, α4, α5, β1 and β5 integrins precisely distribute at the apical surfaces of apposed endometrial LE and chorion along expanses of the interplacentomal endometrial–placental interface between Days 60 and 80 of gestation, suggesting engagement of these integrins with the ECM to stabilize adhesion between endometrial LE and chorion in response to the increasing mechanical stress on this interface by the increasing size of the fetus and volumes of fetal fluids. An advancement is the clear co-localization of OPN and integrins at the endometrial–placental interface throughout gestation in sheep. The comprehensive nature of these results provide evidence that integrins potentially interact with OPN to play key roles in the mechanisms required for implantation and placentation throughout pregnancy in sheep and have implications concerning implantation and placentation in other species.

Effects of Alfaxalone or Propofol on Giant-Breed Dog Neonates Viability During Elective Caesarean Sections

Attention must be paid to C-section anesthesia effects on mother and offspring. Alfaxalone induction results in improved puppy viability when compared to propofol. The present study aims to evaluate effects of alfaxalone or propofol induction for elective C-section on newborns, expressed as Apgar score and fetal fluids cortisol concentration. Anesthesia was induced with alfaxalone 3 mg/kg iv in 5 bitches (group A), and propofol 4 mg/kg iv in another 5 (group P), maintained with isoflurane. Amniotic and allantoic fluids were collected to determine cortisol concentration. Apgar score, litter size, newborn gender, birth-weight, maternal age, and parity were recorded. ANOVA, U Mann-Whitney test and ANCOVA assessed the effects of drugs on the Apgar score and fetal fluids cortisol. Thirty-six puppies were randomly selected for the study: 16 from group A and 20 from group P. Only the Apgar score significantly differed between groups. ANCOVA confirmed a significantly higher Apgar score in group A underlining the influence of fetal fluids cortisol concentrations, both resulting in covariates. Present results confirm the effect of anesthesia on the Apgar score of newborns, which is significantly higher for alfaxalone than propofol, suggesting the use of fetal fluids cortisol as a covariate. These findings could be a starting point for further investigations when less viable puppies are detected or expected, such as during an emergency C-section.

Equine Review

Introduction: This edition of Equine Review has a flavour of equine reproduction and considers papers with practical application. Topics include diffusion of enrofloxacin into fetal fluids and effects on fetal cartilage; the effect of artificial vagina lubricants on sperm quality; the use of slow-release oxytocin for oestrus suppression in mares; and activity of ceftiofur and penicillin with gentamicin in postpartum uterine disease.

Fetal-fluid proteome analyses in late-term healthy pregnant mares and in mares with experimentally induced ascending placentitis

Characterisation of fetal fluids in healthy and disease states of pregnant mares can help to unravel the pathophysiology and to identify putative markers of disease. Thus, this study aimed to compare the protein composition of: (1) amniotic and allantoic fluids of healthy mares obtained immediately after euthanasia and (2) allantoic fluid harvested via centesis before and after experimental induction of placentitis via transcervical inoculation of Streptococcus equi ssp zooepidemicus in healthy mares. Fetal fluids were analysed with a high-throughput proteomic technique after in-gel digestion. Statistical comparisons were performed following normalisation of peptide spectral match. Global normalisation was performed to calculate relative expression. There were 112 unique proteins present in both allantoic and amniotic fluids. There were 13 and 29 proteins defined as amniotic- or allantoic-specific respectively that were present in at least two fluid samples. Another 26 proteins were present in both amniotic and allantoic fluids. Panther DB functional classification grouped fetal-fluid proteins as transfer carriers, signalling molecules, receptors, immunity, hydrolase, enzymes, membrane traffic, cytoskeleton, cell adhesion, calcium binding and extracellular matrix. Experimentally induced placentitis resulted in 10 proteins being upregulated and 10 downregulated in allantoic fluid. Newly identified proteins and changes in the fetal-fluid proteome provide clues about the physiology of pregnancy and pathogenesis of placentitis.