Canine parturition: what is known about the hormonal setting?

Parturition is a challenging physiological process with perfect timing dictated by the events leading to the end of pregnancy in the female, and by the maturation of the fetus(es). The process of parturition remains an intricate interaction of hormones in a fine-tuned timing that remains to be better elucidated in the dog. In the dog pregnancy is maintained by the progesterone production by the corpora lutea, in which some hormones play a luteotrophic action. At term of pregnancy, the pre-parturient luteolytic cascade is the most apparent event, characterized by a rapid decline in plasma progesterone concentrations and useful in predicting the onset of spontaneous parturition. In contrast, cortisol plasma concentrations measurement showed high variability and suggested to be related to the stress condition instead of the onset of parturition. Both prostaglandin F2α and E2 concur in the process of parturition in the dog. The measurement of oxytocin plasma concentrations, also very variable, is implicated in uterine contractions. The measurement of plasma oxytocin concentrations showed to be useful to distinguish between normal and disturbed parturition, allowing the prompt recognition of dystocia and the immediate obstetrical intervention. In contrast to other species, no significant roles of estrogens for the initiating of parturition were demonstrated. Relaxin, the main pregnancy hormonal marker in the dog, beside an endocrine action, is also supposed to play a paracrine/autocrine role at the utero-placental unit and to support the maintenance of high plasma progesterone concentrations through a luteotrophic action, concurring to the main action of prolactin. Although important information has been provided, some aspects in the understanding the hormonal interactions and action timing implications in the process of parturition in the dog remains to be better investigated and represent intriguing topic for basic knowledge and applied research.

Hair cortisol and dehydroepiandrosterone sulfate concentrations in healthy beef calves from birth to 6 months of age.

Cortisol (C) and dehydroepiandrosterone (DHEA) are recognized as the main fetal steroids, and they are likely to influence fetal development and have long-term effects on newborn hypothalamic-pituitary-adrenal axis (HPA) function. DHEA is often measured as its sulfates and expressed as DHEA-S. Hair analysis represents a promising methodological approach for the non-invasive measurement of steroids, allowing for a retrospective analysis of the total exposure to steroids over time, and avoiding the influence of acute events or circadian fluctuations. Hair cortisol and DHEA concentrations have been investigated in cows, but no studies have been performed on calves. The object of this study was to evaluate hair cortisol (HC) and hair DHEA-S (HDHEA-S) concentrations in beef calves from birth to six months of age. Hair samples of 12 beef calves (seven males, five females) were firstly collected at birth (T1) and then every three weeks up to six months of age (T2-T10), collecting only the re-grown hair. HC and HDHEA-S were analyzed by radioimmunoassay (RIA). Calves sex, weight and APGAR score were registered immediately after birth. Statistical analysis revealed that both HC and HDHEA-S were influenced by sampling time (P < 0.001). HC concentrations were higher at T1 compared to all subsequent samplings (T2-T10, P < 0.01); HC concentrations were higher at T2 compared to T4-T10 (P < 0.01), while no further changes were detected from T3 onward. Higher HDHEA-S concentrations were registered at T1, T2 and T3 compared to all the other samplings (P < 0.01). No correlation was found between hair concentrations of both steroids and calf sex or birthweight. APGAR score was negatively correlated only with HC at birth (P < 0.05). These data demonstrate that C and DHEA-S are quantifiable in the hair of calves and are influenced by their age. The higher HC detected at birth (T1) probably reflects the high serum C concentrations present late in pregnancy and increased by the fetal HPA axis, by which parturition is initiated in cows. The highest HDHEA-S at birth (T1) in calves indicates that the largest amounts of DHEA and its sulfates are produced during fetal development. Moreover, the findings of higher HC at three weeks after birth and of higher HDHEA-S until six weeks after birth, suggest that C and DHEA secretion continues also beyond birth, and that these steroids could be involved in the events occurring during the challenging first weeks of age in the calf.

Platelet-rich plasma uterine infusion and pregnancy rate in barren mares with chronic degenerative endometritis.

The chronic degenerative endometritis (CDE) is recognised as directly related to age and infertility in the mare. In this study, the 14 days post ovulation (PO) pregnancy rate was assessed in 60 barren mares affected by CDE, submitted to platelet-rich-plasma intrauterine infusion (PRPI) 24 hours PO. Data showed a significant positive effect of PRPI on the chance to become pregnant in mares affected by Kenney I-II CDE. The overall 14 days PO pregnancy rate was 75%. Sixty-nine % of the overall pregnancies was achieved with the 1st PRPI, but an additional 31% was obtained with the 2nd PRPI at the subsequent heat, increasing significantly the number of pregnancies. Although the 1st PRPI lead to a higher pregnancy rate than the 2nd PRPI (52 vs 23%), the repetition of the treatment had a significant positive effect on the overall pregnancies. Mare's young age positively influenced the chance to become pregnant and was associated to endometritis Kenney I. Mares with endometritis Kenney I had better chance to become pregnant than those with endometritis Kenney II. The PRPI proved, therefore, to be beneficial in barren mares with CDE, and resulted in high overall pregnancy rate achievement in barren mares.

Effect of age and of reproductive status on reproductive indices in horse mares carrying mule pregnancies.

Mules (Equus mulus) are interspecific hybrids derived from crossing horse (Equus caballus) mares with jackasses (Equus asinus). In Italy, the production of mules is very important for the special environmental areas, where they are used as working animals, instead of the forbidden agricultural machines. Although many horse mares carry mule pregnancy successfully at term, low fertility of mares used for producing mules was reported, but limited data about reproductive efficiency in mares carrying mule pregnancy, especially when age and reproductive status are concerned, are available. Therefore, the present study aimed to assess the effect of age and of reproductive status on some reproductive indices of horse mares carrying mule pregnancy, in the final purpose to better clarify factors affecting reproductive performances in the valuable production of mules as working animals under special environmental areas in Italy. The study, performed on 96 draft mares inseminated with Martina Franca donkey semen, showed a satisfactory 89.6% 14 days PO pregnancy rate, lower pregnancy losses (19.8%) than those previously reported for mule pregnancies, leading to satisfactory foaling rates (71.9%), similar to those reported for intraspecific horse pregnancies. A clear, significant detrimental effect of mare's age on pregnancy losses, foaling rate, and on number of estrous cycles needed to achieve pregnancy, was found. Reproductive status also affected the reproductive indices, with significantly higher pregnancy losses and lower foaling rate in barren mares, reopening the question about the possible lower reproduction efficiency in horse mares carrying mule pregnancy. The present study results, although mostly descriptive, add useful information about some factors affecting reproductive performances in the valuable production of mules as working animals under special environmental areas in Italy.

Effect of breed body-size on leptin amniotic fluid concentrations at term pregnancy in dogs.

Because of the need to improve the knowledge about canine perinatology, and given the major role of fetal fluids in sustaining the course of pregnancy and fetal development, an in-depth analysis to better understand the role of some hormones in these compartments is essential. Among all, leptin is recognized to play a key role not only on the energetic homeostasis, but also at multiple levels, influencing the control of reproduction, food assumption and metabolism. Even if in humans and other species it is reported the presence of leptin receptors during fetal development, very little is known about the canine species, in which the role of leptin still needs to be fully understood. The present study aimed to assess the amniotic fluid leptin (AFL) concentrations at term pregnancy in healthy dogs, and to evaluate the possible influence played by breed body-size (after assessment of correlation with maternal bodyweight and placental weight), or other maternal (age, parity, and the so-called "litter effect") and neonatal (gender, birth weight, litter size) parameters on AFL concentrations, analyzed by ELISA test. The study was performed on 90 healthy, viable and normal weighted puppies, 39 small-sized (adult body weight < 10 kg) and 51 large-sized (adult body weight > 25 kg), born by 29 purebred, healthy bitches, submitted to elective Caesarean section because of breed-related or individual high risk for dystocia. The results showed that the mean AFL concentration in the small-sized puppies was significantly (p < 0.05) higher in comparison to large-sized puppies (867.48 vs 698.42 pg/ml), while all the other studied parameters did not show to influence AFL concentrations. In conclusions, the present study showed significant higher at term AFL concentrations in small-sized as compared to large-sized breeds, suggesting an influence of breed body-size on fetal metabolism, as previously reported for NEFA and IGF-I.

Perinatal concentrations of 17ß-estradiol and testosterone in the toe claws of female and male dogs from birth until 60 days of age.

This study was conducted to assess concentrations of 17ß-estradiol (E2) and testosterone (T) in toe claws of puppies collected at birth, at 30 and at 60 days of age, evaluating changes relating to age and effect of puppy sex, Apgar score, bodyweight at birth, "litter effect", litter size, and maternal age. Puppies (n = 89), 46 males and 43 females, with normal weight and without malformations, were assigned for the study. Within 12 h of birth tips of toe claws were clipped, and the re-growth tissue of the claws was collected at 30 and 60 days of age. Steroid quantifications occurred using a radioimmunoassay. The results indicated there were lesser concentrations (P < 0.001) of both hormones at 30 and 60 days of age than at birth and that concentrations were similar at 30-60 days of age. There were greater (P < 0.001) T concentrations in males than females, with there being an interaction between sex and sampling time (P < 0.01). The Apgar score was positively (P < 0.001) related to T concentrations in toe claws at birth. The bodyweight was positively correlated (P < 0.05) with T concentrations, with an interaction among puppy sex, bodyweight and sampling time (P < 0.05). Results of the present study confirmed the usefulness of toe claws as a matrix for study of hormonal changes in perinatology of dogs. Results of the study also indicate there are greater E2 and T concentrations at birth compared with 30 and 60 days of age that could be the result of these prenatal steroids affecting fetal development.

Biochemical composition of fetal fluids in at term, normal developed, healthy, viable dogs and preliminary data from pathologic littermates.

A proper canine neonatal assistance, required to reduce the high perinatal loss rate, imply a full knowledge about the fetal-to-neonatal physiology. Because fetal fluids play an important role throughout mammals pregnancy, influencing fetal growth and development, fetal well being, and contributing to guarantee the most suitable environment for the fetus, the knowledge about fetal fluids biochemical composition is of major importance. At first, the biochemical composition of fetal fluids collected by normal developed, healthy and viable newborns, is necessary to depict the normal features, and represent the first step for the further detection of abnormalities associated to fetal/neonatal distress and useful for the early identification of newborns needing special attention, immediately after birth. The present study was aimed to define the biochemical composition of amniotic and allantoic fluids collected from fetus delivered by caesarean section at term of pregnancy. To reduce the possible confounding effect of maternal labor or troubles at parturition, fetal fluids were collected only from puppies born by elective caesaeran section, at term of normal pregnancies. Fetal fluids from 76 puppies, 70 normal and six pathologic newborns, born by elective caesarean section were collected and analyzed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, lactate dehydrogenase (LDH), creatine-kinase (CK), alkaline phosphatase (ALP), creatinine, urea, amylase, lipase, gamma-glutamyl transferase (γ-GT), triglycerides, cholesterol, total proteins, albumin, globulins, glucose, magnesium, potassium, chloride, sodium, calcium, phosphorus and osmolarity. No significant differences were found between biochemical composition of amniotic or allantoic fluid in normal and pathologic newborns, maybe due to the small number of the pathologic puppies. Although some correlations between the two fluids were found (albumin, phosphorus, glucose and triglycerides), the results showed significant differences between the amniotic and allantoic biochemical composition (for all the parameters, except of alanine aminotransferase, triglycerides, cholesterol, albumin, amylase and glucose), suggesting that diverse sources could concur to the final composition of each fluid. A wide variability within and among litters was found for both amniotic and allantoic biochemical composition, and for some parameters an influence of breed body size (amniotic amylase, cholesterol, and allantoic calcium and glucose), maternal parity (amniotic and allantoic CK, glucose, LDH, chloride) and newborn gender (allantoic phosphorus) was found. Further investigations are needed for addressing the origin of each fetal fluid biochemical composition in the dog and also to indeep possible differences in fetal fluids biochemical composition between normal and pathologic puppies, providing potential markers for the quick identification of newborns that need special surveillance and cares immediately after birth.

Small-sized newborn dogs skeletal development: radiologic, morphometric, and histological findings obtained from spontaneously dead animals.

BACKGROUND: Very little is known about neonatal skeletal development in small-sized purebred dogs. In order to improve this knowledge, 27 spontaneously dead puppies belonging to small-sized breeds were enrolled in this study for radiologic, histological and morphometric investigations. RESULTS: The appearance of the limb secondary ossification centers and the onset of their formation were clearly observed by x rays and confirmed by histological evidences. Radiographic and anatomic measurements of limb bones length and skull length and width were positively correlated with body weight and age of the subjects and the body weight was positively correlated with radius bone mineral density, as demonstrated by dual-energy x-rays absorptiometry. CONCLUSIONS: These data provided original information on the growth of newborn small-sized breed dogs, and suggest that cadavers may be useful to study skeletal development.

Luteal activity and effect of dietary energy restriction on follicular development in lactating cows.

The aim of this research has been to evaluate the presence of anomalies in the ovarian cycle activity during postpartum and to verify whether 72-hr dietary fasting during the dominance phase, the phase before ovulation, might modify the ovarian follicle population. The presence of anomalies in ovarian cycle activity has been evaluated in 30 Italian Friesian cows starting from 20 days postpartum until 211 days of lactation. Long oestrus and brief dioestrus or scarce luteal activity have been the main anomalies found through measuring progesterone concentrations in the whey. Until 100 days of lactation, the BCS values of the problematic animals have been significantly lower than those in animals with normal ovarian activity. After 100 days of lactation, the ovarian anomalies continued to appear despite the fact that all the animals have reached comparable BCS values. Starting from the results of this trial, the effect of 72-hr dietary fasting on dominant follicles has been studied in six cows. Ultrasonography revealed that the diameter of the follicles at 71 days postpartum has been significantly lower than at 181 days. A 72-hr dietary restriction at 101 and 211 days postpartum did not affect the size of the dominant follicle. However, at 101 days postpartum, half of the animals presented follicular cysts. The effect of fasting differed if the animal has been in early postpartum or 211 days of lactation. Further researches are necessary to understand how different metabolic conditions can modify the follicular population but on the other hand the study shows the utility for farmers and field veterinarians of monitoring the resumption of the ovarian cycle postpartum through the whey progesterone concentrations.

Cortisol fetal fluid concentrations in term pregnancy of small-sized purebred dogs and its preliminary relation to first 24 hours survival of newborns.

Along the perinatal period, in mammals, cortisol (C) plays a pivotal role in the final intrauterine fetal maturation and in the early neonatal adaptation. Because of the scarce knowledge about canine perinatology, the present study was aimed to assess the C concentrations in amniotic and allantoic fluids collected, without invasiveness, from small-sized, purebred newborn puppies born by elective cesarean section, at term of pregnancy. Possible correlations between fetal fluid C concentrations and maternal parity, litter size, birth weight, Apgar score, were evaluated. In addition, the possible effect of fetal fluid C concentrations on newborn survival at 24 hours of age, and the effect of the litter or the newborn gender on fetal fluid C concentrations were also assessed. The results, obtained from 50 born alive, normal-weight puppies, without gross physical malformation, showed that C concentration was higher in allantoic than in amniotic fluid (P < 0.01), even if a strong positive correlation between the two fluids C concentration was found (P < 0.0001; R = 0.83). Neither amniotic nor allantoic C concentrations were correlated to maternal parity, litter size, birth weight, and Apgar score. Interestingly, higher amniotic (P < 0.05), but not allantoic, C concentrations were found in puppies not surviving at 24 hours after birth. Therefore, it could be suggested that this parameter may be useful for the recognition, at birth, of puppies needing special surveillance during the first day of age. A significant (P < 0.001) effect of the litter in both amniotic and allantoic C concentrations was found. In conclusion, the present results showed that in small-sized purebred puppies, born at term by elective cesarean section, the exact fetal, maternal, or placental source contributing to fetal fluid C concentrations remains to be clarified. From a clinical perspective, however, the evaluation of amniotic C concentration at birth seems useful for the detection of puppies that need special surveillance during the first 24 hours of age, and should be coupled to the early newborn evaluation by Apgar score. However, the small total number of newborns, and especially of the dead puppies enrolled in the present study, suggests that further, more-focused investigations on a large number of subjects are needed before the method could be considered for application in the clinical practices.

Correlation between some arterial and venous blood gas parameters in healthy newborn Martina Franca donkey foals from birth to 96 hours of age.

In neonatology, blood gas analysis is a useful tool in the evaluation of the health of newborns and plays a key role in early detection of critically ill subjects. Because blood gas analysis parameters have not previously been studied in any depth in donkey foals, this study was performed on 16 healthy Martina Franca donkey foals born after an uncomplicated delivery. Arterial and venous blood samples were collected at 5 minutes and at 12, 24, 72, and 96 hours of age. Blood gas analysis was performed by a portable analyzer, measuring arterial and venous total carbon dioxide, carbon dioxide partial pressure (pCO2), oxygen partial pressure (pO2), oxygen saturation (sO2), bicarbonate, base excess (BE), pH, and lactate (LT). Lower blood pH values, pO2 and sO2, and a higher level of lactate were found at birth in comparison with subsequent sampling times. This moderate acidotic profile disappeared at 12 hours, when all the parameters became constant until the end of the study period. As expected, significant differences between arterial and venous blood gas parameters related to the oxygenation, such as pO2 and sO2, and partially carbon dioxide partial pressure were found, whereas total carbon dioxide, pH, BE, and LT were comparable in arterial and venous blood samples. For these latter parameters, the highly significant correlation between arterial and venous findings suggests that venous samples could be an acceptable alternative to the arterial sample for blood gas analysis in newborn donkey foals, when the oxygenation status of the patient is not the first goal of patient analysis.

Reproductive characteristics of foal heat in female donkeys.

In this study, the first postpartum heat, termed the foal heat, characteristics and performance in female donkey (jenny) of Martina Franca are described. To this end, the follicular development of 42 jennies during foal heat was compared with that of 31 jennies at the third estrus after foaling. Estrus length (7.1±0.9 and 6.8±0.7 days), follicular development and preovulatory follicle size (43.7±3.5 and 45.1±2.5 mm) were similar between jennies during the foal heat and during the third estrus after foaling. The pregnancy rate at day 14 was significantly lower in the foal heat jennies (57.1%) than the third estrus jennies (82.3%). However, the pregnancy rate at day 14 in foal heat jennies increased significantly when the onset of foal heat was ⩾8 days after foaling (93.8%) or when the ovulation happened ⩾12 days after foaling (85.7%). The data provided in the present study suggest that the foal heat in the endangered jennies of Martina Franca could be successfully utilized to reduce the interpartum period if the onset of foal heat occurs >8 days after foaling.

Cartilage canals in newborn dogs: histochemical and immunohistochemical findings.

Cartilage canals (CCs) are microscopic structures involved in secondary ossification centers (SOCs) development. The features of CCs were investigated in the humeral and femoral proximal epiphyses of small-sized newborn dogs (from premature to 28 days after birth) with histochemical and immunohistochemical approaches. Masson's Trichrome revealed a ring-shaped area around CCs, which changes in colour from green (immature collagen) to red (mature collagen) as ossification progresses; perichondrium staining always matched the ring colour. Safranin-O was always negative. Immunohistochemical analysis revealed immunopositivity for both collagen type I and V around the CCs; collagen type II was negative. CCs count showed a tendency to be higher in the humerus than in the femur. This work enlightened for the first time changes in composition of CCs surrounding matrix during SOCs development in dogs, paving the way to further investigations.

Assessment of canine neonatal viability-the Apgar score.

Perinatal mortality is relatively high in dogs, with deaths peaking around the time of birth and during the first week of age. Among the several causes of canine perinatal mortality, whelping is the greatest cause. Therefore, early neonatal assistance at birth should be mandatory with dogs. In comparison with human neonatology, knowledge and technological ability in canine neonatology is tremendously scarce. The Apgar score for the newborn viability assessment at birth represents a feasible method for the prompt recognition of newborns that will need special assistance immediately after birth. The five parameters of the Apgar score were adapted to the canine species by different studies. Advantages and limits, as well as clinical applications, are presented and discussed in further detail. It was concluded that the Apgar score represents the easiest and simplest, non-invasive and reliable method, that could be performed under every clinical and practical condition, for newborn puppies viability evaluations and short-term survival prognosis.

Transrectal ultrasonographic evaluation of combined utero-placental thickness during the last half of pregnancy in Martina Franca donkeys.

In the recent years, the donkey population decreased dramatically so that many breeds are presently considered as endangered. In comparison to the horse, the donkey placenta still remains not completely studied. In the horse, one of the diagnostic tools useful to identify pregnant mares at risk of abortion or premature delivery, include the transrectal ultrasound examination of the uterus and its contents; and especially of the combined thickness of the uterus and of the placenta (CUPT). Since the CUPT was never investigated in donkeys, the present study was aimed to define the transrectal CUPT values during the last half of pregnancy in 20 Martina Franca jennies. Foalings times, foals characteristics and placental gross appearance, and measurements were also evaluated and values resulted always within normality. Differently to the mare, a continuous significant CUPT increase between the sixth to the 12 months of pregnancy, and a substantial increase from the ninth to the 12th month of pregnancy, was found. Although statistically not evaluable, the CUPT values recorded from three jennies with pregnancy loss did not show evidence of CUPT increases. In conclusion, normal CUPT values from the sixth to the 12th month of pregnancy in Martina Franca donkeys are provided, but further investigations are needed to define possible breed or body-size CUPT specific differences, as well as the CUPT values during pregnancy disturbances or placental abnormalities.

Coat and claws as new matrices for noninvasive long-term cortisol assessment in dogs from birth up to 30 days of age.

The last stage of fetal development and the neonatal period represent the most critical phases for the mammals' offspring. In the dog, the knowledge about the final intrauterine fetal development and biology, as well as about the neonatal physiology, remains scarce. Hormonal changes occurring in the last intrauterine fetal phase and during the early neonatal age are still not completely clear, probably because of the invasiveness related to the collection of the more common biological matrix, represented by circulating blood. Toward term of pregnancy, during parturition, and after birth, the hypothalamic-pituitary-adrenal axis is a key system regulating several physiological processes, and its activity was previously investigated by blood analysis, considered an invasive procedure providing a single-point measurement. In respect to animal welfare, and for a more correct long-term retrospective investigation, noninvasive hormonal studies were performed firstly on the hair of humans and coat of animals and, more recently, in the nails of human beings. This study was aimed to assess cortisol (COR) in coat and claws of newborn puppies and to evaluate the possible influence of the newborn gender, breed body size, and age on coat and claws COR concentrations. The results obtained from 165 newborn puppies evidenced that coat and claws COR levels were highly correlated each other (P < 0.0001), although the COR accumulation in the two matrices was different in relation to the class of age. Moreover, the puppies age influenced both coat and claws COR concentrations (P < 0.05), with premature puppies showing higher values when compared to term born-dead puppies or puppies dead between 1 and 30 days of age. The present study reported that COR is quantifiable in coat and claws of newborn dogs. Moreover, both matrices appear as useful tools for new, noninvasive, long-term perinatal and neonatal researches also in canine species.

Endothelin 1 in healthy foals and in foals affected by neonatal diseases.

In newborn babies, endothelin 1 (ET-1), a potent vasoconstrictor, increases during septicemia and severe respiratory syndromes. Because equine neonatal sepsis (ENS) and perinatal asphyxia syndrome (PAS) are major causes of morbidity and mortality in newborn foals and because no information on the concentration of ET-1 in healthy and sick foals has been reported yet, the aims of this study were (1) to define the serum concentration of Big ET-1 in healthy neonatal foals during the first week of age; (2) to preliminarily explore the diagnostic and prognostic role of Big ET-1 during ENS and PAS. Six healthy and 23 sick foals affected by ENS and/or PAS were enrolled in the study. In healthy foals, Big ET-1 concentration increased in the first hours of life until 24 hours after birth, and it remained constant during the first 3 days, then gradually decreased becoming significantly lower from Day 4 onward (P < 0.05). In sick foals, only 26.1% of animals showed higher values of Big ET-1 than controls at admission, and no difference between surviving and nonsurviving foals was found. Because in nonsurviving foals, Big ET-1 remained over the maximum value recorded in clinically healthy horses or, when normal at admission, increased over time; this study suggested that repeated measurement of Big ET-1 during hospitalization may be helpful in monitoring the course of the disease. In conclusion, possible prognostic information may be obtained by repeated analysis of Big ET-1 during hospitalization, but further studies are needed.

Immunoglobulins G and lysozyme concentrations in canine fetal fluids at term of pregnancy.

In the dog, the endotheliochorial placenta allows only the 5% to 10% transfer of maternal antibodies to the fetus, but the timing and the factors influencing the immunoglobulin G (IgG) transplacental transport were not fully investigated. The aims of the present study were the following: (1) to assess the presence of both IgG and lysozyme in amniotic and allantoic fluids collected from fully developed and viable newborn puppies born by elective cesarean section at term and possible correlations between amniotic and allantoic IgG and lysozyme levels; (2) to verify possible differences in IgG and lysozyme concentrations between the two fluids; and (3) to detect possible differences in IgG and lysozyme fetal fluid levels in relation to the maternal breed body size and parity, as well as to the neonatal gender. The study, performed on 41 purebred bitches submitted to elective cesarean section at term, enrolled 142 puppies, 74 males and 68 females, born mature, viable, without gross malformations, and with a normal weight. At surgery, a total of 129 amniotic and 84 allantoic samples were collected for IgG and lysozyme analysis. Class G immunoglobulins and lysozyme were detected in both fluids, but IgG concentrations were higher (P < 0.01) in amniotic fluid. Moreover, a significant positive correlation (P < 0.01) between IgG amniotic and allantoic levels, but not for lysozyme, was observed. A significant effect of the maternal parity (P < 0.05), but not of the breed body size, on the amniotic IgG concentrations was found, whereas the newborn gender was not associated to different IgG or lysozyme amniotic or allantoic levels. Given the significant contributions of fetal fluids to fetal and neonatal health, the results reported that the amniotic and allantoic fluids play a role in the immune protection of the fetus/newborn also in canine species. However, additional research is needed to better elucidate both the origin of IgG and lysozyme and the factors influencing the wide interindividual variations.

Blood analysis in newborn donkeys: hematology, biochemistry, and blood gases analysis.

The knowledge of reference ranges for hematologic, biochemical, and blood gas parameters in the different species and the influence of breed and age on them is a fundamental tool for the clinician. For this reason, the aim of this study was to evaluate the age-related changes of hematologic and biochemical parameters in Martina Franca donkey foals during the first 3 weeks of life and of blood gases during the first 24 hours of age. Fifteen healthy donkey foals were enrolled; blood samples were collected from each foal at 10 minutes after birth, 1 hour after the first and second suckles, 12 and 24 hours after birth, daily from Day 2 to 7, and at Days 10, 14, and 21 of life. Erythrocytes, leukocytes, and platelets counts were assessed; also metabolic (alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, creatinphospokinase, lactate dehydrogenase, alkaline phosphatase, glucose, blood urea nitrogen, creatinine, total proteins, albumins, cholesterol, and total bilirubin) and electrolytic parameters (Ca, P, Mg, Na, K, and Cl) were evaluated. Finally, blood gases and metabolic parameters (pH, pCO2, pO2, sO2, TCO2, HCO3, lactate, and base excess) on venous blood were assessed with a portable analyzer. A statistical analysis to evaluate the influence of age and sex was performed. Several differences were found between sampling times, demonstrating that age influences these parameters. Moreover differences were found compared with data reported in literature for donkey foals of another species, horse foals, and adult donkeys. Although a great interindividual variation for some parameters exists, this study demonstrated that interval references should be addressed not only to different species, but also to specific breeds and to the neonatal period.

Effect of the season on some aspects of the estrous cycle in Martina Franca donkey.

The Martina Franca (MF) donkey breed, with 48 jackasses and 515 jennies, is considered an endangered breed according to the data from the Monitoring Institute for Rare Breeds and Seeds in Europe. The knowledge of the estrous cycle characteristics has a great impact for assisted reproduction, especially in endangered species. In this study, the estrous cycle characteristics were investigated in 12 MF jennies throughout the year. Estrous cycle, estrous and diestrous lengths, follicular growth and ovulation, and estradiol-17ß (E2) and progesterone (P4) plasma concentrations were monitored in MF jennies and compared in different seasons. In all jennies (100%) estrous cycle was detected during the whole year, with no differences in the estrous cycle length among seasons. However, a significant increase of estrous length in spring and summer compared with autumn and winter was found. Diestrus was shorter in summer than in the other seasons. Estrous behavior was always shown and characterized by rhythmic eversion of the vulvar labia (winking) with exhibition of the clitoris, urination, male receptivity and clapping, with sialorrhoea, neck and head extension, and back ears. Estrus was characterized by the ovulation of a larger follicle in spring and summer than in autumn and winter. The pattern of E2 and P4 plasma concentrations during the estrous cycle were similar to that reported for the mare, but without differences among the four seasons, so that a negligible effect of environmental conditions on ovarian E2 and P4 secretion was hypothesized, despite the larger diameter of the ovulating follicle in spring and summer.