Perinatal cortisol and blood glucose concentrations in bitches and neonatal puppies: effects of mode of whelping.

The decision on how and when to assist whelping is crucial for the survival rate of puppies and health status of the dam. However, medical or surgical therapy in dystocia can impact both maternal and neonatal stress and glucose response differently. This study aims to compare perinatal cortisol and glucose among different modes of delivery in bitches and neonates. We analyzed 50 puppies derived from 27 healthy bitches. According to the condition at birth, bitches and their puppies were allocated into either a Eutocia Group (vaginal birth with no whelping assistance), Fetal Dystocia Group (whelping assistance with fetal manipulation), Maternal Dystocia Group (whelping assistance because of partial uterine inertia corrected by oxytocin administration), or Cesarean Section Group (fetal or maternal dystocia bitches subjected to C-section). Maternal blood cortisol and glucose concentrations were analyzed during the perinatal period (prepartum, intrapartum, postpartum, and 1 h after postpartum). Neonatal blood samples were collected within 5 min and 1 h after birth for assessment of cortisol and glucose. Maternal dystocia bitches had higher cortisol concentrations at postpartum than the Fetal Dystocia Group. At 1 h postpartum, the Cesarean Section Group had higher cortisol concentrations compared with fetal dystocia bitches. The Eutocia Group presented increased cortisol concentrations at intrapartum and postpartum, whereas fetal dystocia bitches had higher intrapartum cortisol concentrations than at 1 h postpartum. The Maternal Dystocia Group presented higher postpartum cortisol concentrations than at prepartum and 1 h postpartum. Maternal glucose had a progressive increase throughout peripartum and was higher during postpartum and at 1 h postpartum. C-section bitches had the highest blood glucose concentration. Neonatal cortisol concentrations at birth were higher than 1 h after birth. Fetal dystocia puppies had higher cortisol concentrations, whereas caesarian section puppies had lower cortisol levels. Fetal dystocia and C-section puppies had higher glucose concentrations than the Eutocia Group. In conclusion, maternal dystocia leads to high cortisol concentrations in bitches immediately postpartum, whereas only fetal dystocia causes increased neonatal cortisol concentrations. Moreover, fetal dystocia and C-section are hyperglycemic obstetrical conditions for neonatal puppies; on the other hand, only C-section causes hyperglycemia in bitches.

Morphometric and functional pulmonary changes of premature neonatal puppies after antenatal corticoid therapy.

Among prematurity complications, the most important disorder is structural immaturity and inadequate production of pulmonary surfactant. Betamethasone is the drug of choice to artificially improve pulmonary capacity, thus we aimed to verify the effect of prenatal maternal treatment on lung development of premature puppies. Pregnant bitches were allocated in Term Group (n = 7), Preterm-Treated Group (interrupted pregnancies with maternal administration of betamethasone; n = 7), Preterm-Control Group (untreated interrupted pregnancies; n = 7), Extremely-Preterm Group (interrupted pregnancies at 55d; n = 6). Puppies were subjected to chest radiographic at birth, morphometric description of pulmonary structures and immunohistochemical analysis of surfactant protein B, proliferating cell nuclear antigen and cytokeratin were performed. In Preterm-Treated Group it was possible to more clearly identify cardiac silhouette and lung parenchyma by X-Ray. Saccular formation was higher in Preterm Groups, while Term Group had higher subsaccular development. Lung septation was higher in Treated and Term Groups. Term Group had higher number of cells marked for SP-B, whereas higher proliferation was observed in Extreme-Preterm and Preterm-Control Groups. Preterm Treated and Term Groups had higher tissue differentiation. In conclusion, antenatal maternal corticotherapy in dogs acted by increasing lung morphology and development of areas of gas exchange, regulate metabolism of pulmonary fluids rather than stimulate surfactant production.

Influence of prenatal maternal corticosteroid therapy on clinical and metabolic features and pulmonary function of preterm newborn puppies.

Although the effects of antenatal corticosteroid therapy in clinical improvement and pulmonary maturation in preterm have been described, little is known on premature newborn puppies. This study aimed to evaluate the effect of maternal administration of a single dose of prenatal betamethasone on lung function of preterm newborn puppies in the first hours of life, especially from the clinical point of view and acid-base balance. A prospective study was conducted involving 21 puppies allocated into three experimental groups: Term Group (63 days post-ovulation), Preterm-Treated Group (57 days post-ovulation and maternal administration of a single dose of 0.5 mg/kg of betamethasone) and Preterm-Control Group (57 days post-ovulation). Puppies were analyzed clinically through the Apgar score, heart rate, respiratory rate and neurological tests (muscular tone and irritability reflex) and for oximetry and blood acid-base balance in distinct experimental moments. Premature puppies had marked degree of prematurity, reversed by maternal administration of betamethasone. Prenatal corticosteroid therapy promoted better pulmonary and metabolic condition, with more efficient compensatory response to acid-base imbalance and better pulmonary gas exchange capacity. Therefore, prenatal treatment with betamethasone can be adopted as clinical lung maturation protocol for pregnancies at risk in order to prevent low vitality and increase neonatal survival.

Cytoplasmic droplet acting as a mitochondrial modulator during sperm maturation in dogs.

Motility acquisition during sperm maturation and passage through the epididymis is closely related to mitochondrial function and appears to occur in parallel with cytoplasmic droplet (CD) migration. However, such mechanism remains unclear in dogs. Thus, the aim of this study was to characterize the influence of sperm CD in the mitochondrial functionality during epididymal sperm maturation in dogs. Twenty-one adult dogs were submitted to elective bilateral orchiectomy. Testicles were stored for 18-24h at 5°C and epididymal sperm samples were then collected from different segments of the epididymis (caput, corpus and cauda). Samples were evaluated for computer-assisted motility analysis (CASA), presence of CD (eosin/nigrosin stain), ultrastructural CD analysis and sperm mitochondrial activity (3,3' diaminobenzidine technique) and membrane potential (JC-1 probe). Samples collected from the corpus epididymis showed higher motility and mitochondrial activity in comparison to the caput sperm. Moreover, corpus sperm had lower percentage of proximal droplets compared to caput samples, while mitochondrial membrane potential remained unchanged. Cauda samples showed higher motility, mitochondrial activity and potential, however, lower presence of sperm droplets (proximal and distal). In conclusion, the CD is essential for epididymal sperm maturation in dogs, showing important functions along the transit in the epididymis. In the corpus segment, the migration of the CD along the sperm midpiece provides a high mitochondrial activity and the onset of sperm motility. On the other hand, sperm from cauda epididymis lack CD but suffered lipid membrane changes which allow a maximum mitochondrial membrane potential and motility.

Lipid composition of the canine sperm plasma membrane as markers of sperm motility.

The fatty acid composition of the sperm membrane is an important factor involved in the overall sperm quality, including motility. However, in the canine species, the exact composition of the plasma membrane is still unknown. Therefore, the purpose of this study was to evaluate the plasma membrane lipid composition of motile sperm cells and to compare it with asthenospermic samples, as an attempt to determine possible involvements of membrane lipids in dog sperm cell motility. The sperm-rich fraction of ten mature dogs was collected, and samples were subjected to density gradient centrifugation by Percoll® , in order to separate motile and asthenospermic samples. Processed semen samples were evaluated for sperm motility, plasma and acrosome membrane integrity, mitochondrial activity and susceptibility to oxidative stress. Lipid plasma membrane composition was identified by mass spectrometry (MALDI-MS). The motile sperm samples presented the following phospholipids in a high frequency in the plasma membrane: phosphatidylcholine 38:4 (composed of stearic and arachidonic fatty acids), phosphatidylcholine 36:1 (stearic and oleic fatty acids), phosphatidylethanolamine 34:4 (myristic and arachidonic fatty acids), glycerophosphatidic acid 36:4 (palmitic and arachidonic fatty acids), phosphatidylcholine 40:4 plasmanyl and phosphatidylcholine 40:5 plasmenyl. Furthermore, no lipid markers were found in the asthenospermic samples. Results also indicate that differences on plasma membrane composition between motile and asthenospermic samples are crucial factors for determining sperm motility, sperm functionality and susceptibility to oxidative stress. In conclusion, plasma membrane lipid composition varies considerable between motile and asthenospermic samples. Therefore, lipid markers of sperm motility can be considered, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylcholine plasmanyl, phosphatidylcholine plasmenyl and phosphatidic acid.

The influence of canine brucellosis on morphofunctional features of epididymal spermatozoa: case report / Influência da brucelose canina nos aspectos morfofuncionais de espermatozoides epididimários: relato de caso

The present work reports a clinical case of a mongrel dog, with serological diagnosis of brucellosis, from which epididymal sperm analysis was performed. Sperm samples were collected from different segments of the epididymis (tail, corpus, and caput). Sperm samples were evaluated for computer-assisted motility analysis (CASA), spermatic morphology, mitochondrial activity and sperm plasmatic membrane and acrosomal integrity. Changes in sperm movement patterns were found (progressive motility, percentage of rapid sperm, percentage of rapid velocity, average pathway, curvilinear velocity, velocity straight line, amplitude of lateral head displacement, straightness and linearity), increase of total morphological defects (51%) and absence of sperm mitochondrial activity (20%) were verified, especially for cauda epididymides. We highlight that such changes can contribute to clinical diagnosis of Brucellosis in dogs and to the use of epididymal sperm in reproductive biotechnologies.(AU)

Relata-se o caso de um cão mestiço, com diagnóstico sorológico para brucelose canina, a partir do qual foram realizadas análises do sêmen epididimário. As amostras espermáticas foram coletadas dos diferentes segmentos epididimários (cabeça, corpo e cauda). Foram realizadas as avaliações de motilidade computadorizada do sêmen (CASA), morfologia espermática, atividade mitocondrial, integridade das membranas plasmática e acrossomal. Houve alteração no padrão de movimentação espermática (motilidade progressiva, espermatozoides rápidos, velocidade média da trajetória, velocidade curvilínea, velocidade linear progressiva, amplitude de deslocamento lateral da cabeça, retilinearidade e linearidade), aumento do total de defeitos morfológicos (51%) e da ausência de atividade mitocondrial espermática (20%) dos espermatozoides, especialmente da cauda do epidídimo. Ressalta-se que tais achados podem contribuir para o diagnóstico clínico da brucelose canina e para a utilização do sêmen epididimário em biotecnologias da reprodução.(AU)

Effect of reduced glutathione (GSH) in canine sperm cryopreservation: In vitro and in vivo evaluation.

The aim of this study was to compare the in vitro and in vivo efficiency of different concentrations (0, 10 and 20 mM) of reduced glutathione supplemented to the extender for canine semen cryopreservation. Six normospermic dogs were used and each ejaculate was divided in 3 experimental groups, according to GSH concentration (GSH-0, GSH-10 and GSH-20 Groups). After thawing, samples were evaluated by sperm motility by computer-assisted sperm analysis (CASA), flow cytometric evaluation of plasma and acrosome membrane integrity, mitochondrial membrane potential and activity, chromatin susceptibility to acid-induced denaturation, and measurement of spontaneous and induced production of thiobarbituric acid reactive substances (TBARS). In vivo tests were carried out with GSH-0 and GSH-10 groups, for which six bitches were inseminated with semen cryopreserved in extender without GSH or containing 10 mM GSH. Intrauterine insemination was performed by cervical catheterization on the 5th and 6th days after the LH surge, detected by serum progesterone and LH assays. In the CASA evaluation, GSH-20 group had the lowest total and progressive motility and lower percentage of sperm with rapid and slow speed. Groups treated with glutathione showed lower percentage of acrosome damage, but higher percentage of plasma membrane injury. GSH-20 group had higher percentage of sperm with low mitochondrial activity and higher concentration of induced TBARS. Both groups (GSH-0 and GSH-10) had positive pregnancies. In conclusion, 20 mM GSH supplementation to canine cryopreservation extender promoted sperm damage, especially to mitochondrial activity. However, addition of 10 mM GSH resulted in acrosome protection, preserving fertility rate.

Oxidative stress at different stages of two-step semen cryopreservation procedures in dogs.

Sperm cryopreservation generates sperm damage and reduced fertilization capacity as a consequence of reactive oxygen species formation. Identifying the critical points of the process will contribute to the development of strategies for oxidative stress prevention. Therefore, the aim of this experiment was to verify the occurrence of oxidative stress during the two-step cryopreservation process in dogs. Six healthy mature dogs were used and submitted to the two-step sperm cryopreservation protocol. The sperm analysis was done at three time points: after refrigeration, after glycerolization, and after thawing by sperm motility, measurement of spontaneous and induced oxidative stress, sperm mitochondrial activity, plasma membrane integrity, flow cytometric evaluation of plasma and acrosome membrane integrity, mitochondrial membrane potential, and sperm chromatin structure assay. There was an increase in free radical production after glycerolization (87.4 ± 15.5 ng/mL of spontaneous thiobarbituric acid reactive substances (TBARS) after refrigeration and 1226.3 ± 256.0 ng/mL after glycerolization; P < 0.05), in association with loss of sperm mitochondrial activity. However, frozen-thawed samples had lower sperm motility, lower resistance to oxidative stress (448.7 ± 23.6 ng/mL of induced TBARS after glycerolization and 609.4 ± 35.9 ng/mL after thawing; P < 0.05) and increased lipid peroxidation (4815.2 ± 335.4 ng/mL of spontaneous TBARS after thawing; P < 0.05) as well as increased damage to plasma and acrosomal membranes, compared with refrigeration and glycerolization. In conclusion, the production of free radicals by sperm cells begins during glycerolization. However, sperm oxidative damage intensifies after thawing. Despite intracellular ice formation during cryopreservation, the increased production of reactive oxygen species can be the explanation of the decrease in sperm motility, reduced mitochondrial activity, and sperm plasma membrane and acrosomal damage.

Effects of intratesticular administration of zinc gluconate and dimethyl sulfoxide on clinical, endocrinological, and reproductive parameters in dogs.

Nonsurgical sterilization methods are considered alternative tools for the worldwide challenge represented by canine overpopulation control. Intratesticular injection of zinc gluconate associated with DMSO arises as an option because of the effortless diffusion throughout the testicular parenchyma. This study aimed to verify the effectiveness of a double testicular injection of zinc gluconate associated with DMSO as a chemical contraceptive for male dogs. The study was conducted with 22 dogs treated with two intratesticular injections of the chemical solution (treated group; n = 15) or 0.9% NaCl solution (control group; n = 7) on a monthly interval. All animals were submitted to clinical examination, breeding soundness evaluation including morphologic and sonographic examination of the testes, assessment of libido, volume of the sperm-rich fraction, sperm motility, total sperm count, plasma membrane integrity, sperm morphologic abnormalities, and the total number of morphologically normal and motile sperm in the ejaculate. Blood samples were collected for serum testosterone analysis, and testicular tissue was morphologically and histologically evaluated. No clinical alterations and signs of pain or local sensitivity along the experimental period were noticed. However, the injection of zinc gluconate and DMSO significantly reduced libido and testosterone concentrations (even beyond the reference range for intact male dogs). Impairment of sperm quality-related variables was observed 15 days after the first intratesticular administration of zinc gluconate and DMSO (i.e., decrease in sperm count and sperm motility and an increase in major sperm defects and by this a decrease in the total number of morphologically normal and motile sperm). Testicular ultrasonographic analysis revealed reduction of testicular volume and changes of testicular echotexture in treated animals, compatible with tissue degeneration, fibrosis, and calcification of testicular parenchyma on histologic examination. In conclusion, intratesticular administration of zinc gluconate associated with DMSO reduces reproductive potential which may lead to subfertility or infertility in dogs.

Gene expression of estrogen and oxytocin receptors in the uterus of pregnant and parturient bitches

In the canine species, the precise mechanisms of pregnancy maintenance and the initiation of parturition are not completely understood. The expression of genes encoding the receptors for estrogen (ERα mRNA) and oxytocin (OTR mRNA) was studied in the endometrium and myometrium during pregnancy and parturition in dogs. Real-time PCR was performed to quantify the levels of ERα mRNA and OTR mRNA in the uterus of bitches during early (up to 20 days of gestation), mid (20 to 40 days) and late pregnancy (41 to 60 days), and parturition (first stage of labor). All tissues expressed ERα and OTR mRNA, and are thus possibly able to respond to eventual estrogen and oxytocin hormonal stimuli. No statistically significant differences in the expression of ERα mRNA were verified in the endometrium and myometrium throughout pregnancy and parturition, but expression of OTR mRNA increased at both parturition and late pregnancy. We concluded that the increase of endometrial and myometrial OTR mRNA expression in dogs is not an event dependent on estrogenic stimulation. Moreover, the contractility response of the canine uterus to oxytocin begins during pregnancy and maintains myometrial activity. The expression of OTR mRNA in canine uterine tissues varied over time, which supports an interpretation that the sensitivity and response to hormone therapy varies during the course of pregnancy and labor. Further studies are needed to elucidate the factors underlying the synthesis of uterine oxytocin receptors and the possible role of ERβ rather than ERα in the uterine tissues during pregnancy and parturition in dogs.

Gene expression of estrogen and oxytocin receptors in the uterus of pregnant and parturient bitches.

In the canine species, the precise mechanisms of pregnancy maintenance and the initiation of parturition are not completely understood. The expression of genes encoding the receptors for estrogen (ERα mRNA) and oxytocin (OTR mRNA) was studied in the endometrium and myometrium during pregnancy and parturition in dogs. Real-time PCR was performed to quantify the levels of ERα mRNA and OTR mRNA in the uterus of bitches during early (up to 20 days of gestation), mid (20 to 40 days) and late pregnancy (41 to 60 days), and parturition (first stage of labor). All tissues expressed ERα and OTR mRNA, and are thus possibly able to respond to eventual estrogen and oxytocin hormonal stimuli. No statistically significant differences in the expression of ERα mRNA were verified in the endometrium and myometrium throughout pregnancy and parturition, but expression of OTR mRNA increased at both parturition and late pregnancy. We concluded that the increase of endometrial and myometrial OTR mRNA expression in dogs is not an event dependent on estrogenic stimulation. Moreover, the contractility response of the canine uterus to oxytocin begins during pregnancy and maintains myometrial activity. The expression of OTR mRNA in canine uterine tissues varied over time, which supports an interpretation that the sensitivity and response to hormone therapy varies during the course of pregnancy and labor. Further studies are needed to elucidate the factors underlying the synthesis of uterine oxytocin receptors and the possible role of ERß rather than ERα in the uterine tissues during pregnancy and parturition in dogs.

Association between birth conditions and glucose and cortisol profiles of periparturient dairy cows and neonatal calves.

Parturition in cattle is a stressful event for both the dam and the offspring. Stress and pain can alter the energy profile of calves and calving cows, producing a metabolic imbalance at birth. This study aimed to assess the effects of dystocia and oxytocin and calcium infusion on metabolic homeostasis in dairy cows and calves. Thirty Holstein cows and their calves were divided into three groups: an eutocia group (n=10), in which no calving assistance was needed; a dystocia group, which required mild-to-severe obstetric assistance (n=10); and a uterine inertia group, which was treated with oxytocin and calcium (n=10). To assess serum cortisol and blood glucose levels, blood samples were collected during the peripartum period from cows and during the first hour since birth from calves. All groups were hyperglycaemic following parturition. Infusion of oxytocin and calcium resulted in lower maternal glucose concentrations and lower levels of stress than in cows in the dystocia group. Birth condition was significantly associated with blood glucose and cortisol concentrations in calves. Glucose concentration was lower in calves born with oxytocin and calcium infusion than those born with fetal extraction. In conclusion, assisted calving with fetal extraction causes important metabolic changes for the dam and calf. Conversely, the practice of oxytocin and calcium infusion for hypotonic cows has no harmful effects on metabolic balance and can be safely employed as a medical treatment.

Effect of dystocia and treatment with oxytocin on neonatal calf vitality and acid-base, electrolyte and haematological status.

Under adverse obstetrical conditions, appropriate supervision and assistance during the immediate neonatal period are of the utmost importance, especially for weak calves. The aim of this study was to establish the effects of dystocia and oxytocin infusion on neonatal vitality, acid-base balance, and electrolyte and haematological homeostasis of dairy calves. Data were collected for 30 Holstein calves which were allocated to three groups: normal calving (n = 10); dystocia with mild to severe obstetric assistance (n = 10); and uterine inertia treated with oxytocin (n = 10). All 30 calves exhibited normothermia at birth, but had a significant decrease in body temperature after 60 min. Dystocic calves had lower Apgar scores than calves in the other two groups, and had respiratory and metabolic acidosis. Calves from normal calvings had normal blood pH, but base excess below the reference range. The mean partial pressure (Pa) of oxygen of calves whose dam had been treated with oxytocin was lower than that of calves from normal calvings. In all experimental groups, there was improvement in metabolic status in the first 60 min postpartum as PaCO2 values significantly decreased. All calves had normonatraemia, normokalaemia and normochloridaemia during the study period, but calves born to dams treated with oxytocin had a higher sodium concentration than those in the two other groups.

The oxidative stress, antioxidant profile and acid-base status in preterm and term canine neonates.

During the initiation of neonatal pulmonary respiration, there is an exponential increase in reactive oxygen species that must be scavenged by antioxidant defences. However, neonate and preterm newborns are known to possess immature antioxidant mechanisms to neutralize these toxic effects. The purposes of this study were to compare the development of antioxidant system between preterm and term canine neonates and to evaluate the magnitude of acid-base balance during the initial 4 h of life. A prospective study was conducted involving 18 neonatal puppies assigned to Term Group (63 days of gestation; n = 5), Preterm-57 Group (57 days of gestation; n = 8) and Preterm-55 Group (55 days of gestation; n = 5). Neonates were physically examined through Apgar score and venous haemogasometry within 5 min, 2 and 4 h after birth. No difference on amniotic fluid and serum superoxide dismutase (SOD), glutathione peroxidase (GPx) and the marker of oxidative stress (thiobarbituric acid reactive substances; TBARS) was verified. Irrespective of prematurity, all neonates presented low vitality, hypothermia, acidosis, hypoxaemia and hypercapnia at birth. However, term puppies clinically evolved more rapidly than preterm newborns. During the course of the study, premature neonates presented more severe complications, such as prolonged hypoxaemia and even death. In conclusion, premature puppies have no signs of immature enzymatic mechanisms for controlling oxidative stress, although SOD and GPx may participate in achieving acid-base balance. Aside from initial unremarkable symptoms, premature puppies should be carefully followed up, as they are at high risk of succumbing to odds of prematurity.

Role of residual cytoplasm on oxidative status during sperm maturation in dogs.

During maturation sperm cells acquire their fertilizing ability; however, this event also produces reactive oxygen species and as such local antioxidant protection is required. Thirteen epididymis from dogs were used, and sperm samples were collected from different segments of the epididymis (i.e. caput, corpus and cauda). The samples were evaluated for motility and vigor, permeability of plasma membrane, presence of cytoplasmic droplet, acrosome integrity and the activity of the antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD). Samples collected from the cauda showed higher motility, vigor and lower permeability of plasma and acrosomal membranes in relation to the corpus and caput, indicating different levels of maturity. The enzyme activity remained unchanged in the samples. A significant positive correlation was observed in the epididymal caput, between distal cytoplasmic droplet and SOD, and a negative correlation, in the epididymal cauda, between proximal cytoplasmic droplet and GPx, indicating the physiological need of the specific antioxidants in these segments.

Sperm maturation in dogs: sperm profile and enzymatic antioxidant status in ejaculated and epididymal spermatozoa.

Spermatozoa become more susceptible to the attack of reactive oxygen species during maturation. To avoid oxidative damage, the epididymis must provide the necessary antioxidant protection. The aim of this study was to compare the canine sperm profile and the enzymatic antioxidant status of the ejaculated fractions and samples collected from the different segments of the epididymis (caput, corpus and cauda). Five adult dogs were used, and after 1-3 weeks, subsequently to bilateral orchiectomy and epididymal storage, sperm samples were collected from the different segments of the epididymis. Samples were evaluated for conventional microscopy and computer-assisted motility analysis: sperm plasma membrane permeability and the activity of the antioxidant enzymes catalase, glutathione peroxidase (GPx) and superoxide dismutase (SOD). Samples collected from the caput and corpus showed lower values for most of the motility variables evaluated, indicating different levels of immaturity. Catalase activity was observed only in ejaculated samples. Conversely, GPx activity was higher in the cauda epididymidis. Correlations were found between SOD and GPx and SOD and sperm motility in the epididymal cauda and corpus, highlighting the importance of the enzymes for the protection of spermatozoa during the transit along the epididymis.

Prenatal and neonatal adaptations with a focus on the respiratory system.

Among the modifications that occur during the neonatal period, pulmonary development is the most critical. The neonate's lungs must be able to perform adequate gas exchange, which was previously accomplished by the placenta. Neonatal respiratory distress syndrome is defined as insufficient surfactant production or pulmonary structural immaturity and is specifically relevant to preterm newborns. Prenatal maternal betamethasone treatment of bitches at 55 days of gestation leads to structural changes in the neonatal lung parenchyma and consequently an improvement in the preterm neonatal respiratory condition, but not to an increase in pulmonary surfactant production. Parturition represents an important challenge to neonatal adaptation, as the uterine and abdominal contractions during labour provoke intermittent hypoxia. Immediately after birth, puppies present venous mixed acidosis (low blood pH and high dioxide carbon saturation) and low but satisfactory Apgar scores. Thus, the combination of physiological hypoxia during birth and the initial effort of filling the pulmonary alveoli with oxygen results in anaerobiosis. As a neonatal adaptation follow-up, the Apgar analysis indicates a tachypnoea response after 1 h of life, which leads to a shift in the blood acid-base status to metabolic acidosis. One hour is sufficient for canine neonates to achieve an ideal Apgar score; however, a haemogasometric imbalance persists. Dystocia promotes a long-lasting bradycardia effect, slows down Apgar score progression and aggravates metabolic acidosis and stress. The latest data reinforce the need to accurately intervene during canine parturition and offer adequate medical treatment to puppies that underwent a pathological labour.

Cortisol profile and clinical evaluation of canine neonates exposed antenatally to maternal corticosteroid treatment.

The effects of glucocorticoids on both foetal canine lung and endogenous serum cortisol concentration have not been clearly delineated. Therefore, we aimed to investigate whether maternal corticosteroid treatment can alter maternal and neonatal cortisol profile and improve neonatal vitality. We allocated six bitches of different breeds and their neonates into two groups: control group (CONT)--maternal administration of saline solution at 55 days post-ovulation (n = 3); and betamethasone group (BETA)--administration of a single dose of 0.5 mg/kg betamethasone (Celestone Soluspan(®) ) at 55 days post-ovulation (n = 3). Caesarean sections were scheduled for day 63 after ovulation. However, BETA group dams showed precocious signs of labour, and c-sections were performed at 58 days post-ovulation. Maternal and neonatal evaluations were performed periodically between betamethasone administration and birth, respectively. Neonates from both groups presented unsatisfactory (<5) Apgar score at birth. However, in spite of an earlier improvement on vitality found on CONT group and the premature delivery on BETA group, both groups showed acceptable Apgar score 120 min after birth. Neonatal cortisol concentrations were higher on CONT group compared to BETA group at birth. In addition, a gradual decrease on maternal cortisol concentrations was observed in the BETA group from treatment until parturition. These findings suggest that despite the down-regulation on the hypothalamic-pituitary-adrenal axis and the induction of premature delivery, betamethasone treatment was able to provide similar vitality when compared to the untreated neonates born at term.

Peripartum haemodynamic status of bitches with normal birth or dystocia.

There has been limited investigation of parturition in the bitch and there is little information published on clinical and obstetrical examination other than opinion and anecdote. While there are substantial data on haemodynamic and vascular changes during normal parturition in humans, little is known about the physiological events in the dog. This study was aimed at maternal haemodynamic changes occurring during normal parturition and to investigate how these were modified in bitches with dystocia (DYST) treated either medically or via assisted delivery and caesarean operation. Three groups of 10 bitches were investigated; those with normal parturition, those with DYST corrected by manipulative assistance or caesarean operation and those with uterine inertia treated by oxytocin administration. Heart rate, systolic and diastolic blood pressure, electrocardiogram and blood glucose concentration were measured pre-partum, intra-partum, immediately after parturition and 1 h later. Heart rate was high at all times throughout the study and the majority of bitches had normal sinus rhythm. Blood pressure was generally within the normal range, and although systolic and diastolic blood pressure was highest during the intra-partum period and sometimes during the immediate post-partum period, there were no significant differences between groups. All bitches had blood glucose concentrations within the normal range throughout the study although pre-partum concentrations were statistically lower than many of the other time periods. The study provides useful physiological data that will facilitate monitoring and clinical management of bitches throughout normal parturition and DYST.

Neonatal clinical evaluation, blood gas and radiographic assessment after normal birth, vaginal dystocia or caesarean section in dogs.

This study aimed to standardize signs and diagnostic criteria of respiratory function in newborn puppies delivered normally or after dystocia and caesarean operation. A total of 48 neonates were allocated into groups: eutocia (n = 20), dystocia (n = 8), caesarean (c)-section (n = 20). Neonatal health was assessed using the Apgar score and body temperature was determined at 0, 5 and 60 min after delivery. Venous blood gases (pO(2) and SO(2)) was measured immediately and 60 min after delivery, and a thoracic radiograph was made between 0 and 5 min of life. The c-section group had significantly lower Apgar scores at birth and 5 min. Hypothermia was present at 5 min in the eutocia and c-section groups, and at 60 min in all groups. The eutocia group had an irregular respiratory pattern in 78% of puppies at birth, 27.7% at 5 min and 21% at 60 min compared with 87.5%, 62.5% and 12.5% of the pups in the dystocia group where there was irregular respiratory rhythm, moderate to intense respiratory sounds with agonic episodes. The c-section group had respiratory alterations in 70%, 45% and 16% of puppies at 0, 5 and 60 min, respectively. Radiographic abnormalities were present in 17% of the pups in the eutocia group, 25% of the pups in the dystocia group and 30% of the pups in the c-section group, respectively. The c-section group had significantly lower SO(2) values at 60 min than at birth. All puppies had hypoxaemia, but a significant decrease was observed in the c-section group. Newborn puppies had tissue hypoxia and irregular respiratory pattern at birth. Caesarean-section puppies had lower vitality; however, all developed satisfactory Apgar scores at 5 min of life, regardless of the obstetric condition.