Comparative study on 15-ketodihydro-PGF(2α) plasma concentrations in newborn horses, donkeys and calves.

The aim of this study was to compare the plasma profiles of 15-ketodihydro-PGF(2α) (PGM) in healthy neonates of three different species from birth until the third week of life. Twenty-four horse foals, 12 donkey foals, and 9 calves were studied. Blood samples were collected at 10, 20 and 30 min after birth, at 3, 24 and 72 h after birth, and at 7, 10, 17 and 21 days of life. All mothers experienced normal gestation lengths and normal, spontaneous deliveries. All newborns were judged mature and viable. Hormone concentrations were higher (p < 0.05) in horse foals 20 and 30 min after birth compared to later samples, and at 10 min in donkey foals compared to later samples (p < 0.05). In calves, higher (p < 0.05) concentrations of PGM were observed 10, 20, 30 min and 3 hours from parturition compared to later samples. These findings may be related to increased fetal placental unit production during parturition, while the relatively high PGF(2α) levels in the days after parturition may be connected with their role in completing organ maturation. Despite the existing differences between these species, the statistical analysis did not discover significant differences in PGM profiles during the first 3 weeks of life in donkey, horse and cattle newborns. The low levels observed 10 days after birth are possibly due to a fast completion of maturational development in these species.

15-Ketodihydro-PGF2alpha and cortisol plasma concentrations in newborn foals after spontaneous or oxytocin-induced parturition.

Hormonal changes during early neonatal life play a major role in the physiological processes underlying the maturation of several organs. Since prostaglandins and cortisol are associated with fetal organ system maturation, the aim of this study was to evaluate 15-ketodihydro-PGF(2alpha) (PGM) and cortisol plasma concentrations during the first 21 days after birth in foals born by either spontaneous (24 foals) or low-dose oxytocin (OT)-induced parturition performed after at least 320 gestational days (25 foals) since induction is often considered to be a cause of prematurity. After spontaneous birth, the PGM concentration was significantly (P<0.05) higher at 20 and 30min compared to samples taken several hours or days later, while induced foals showed significantly (P<0.05) higher concentrations at 10, 20, and 30min. Regarding differences between the two groups, the plasma concentration of PGM was significantly higher 10 (P<0.01), 20 (P<0.05), and 30 (P<0.05)min and 3h (P<0.05) after birth in induced foals compared to foals born by spontaneous parturition. It is difficult to determine whether the higher initial PGM concentrations in induced foals is related to higher uterine or fetal PGM release induced by exogenous OT stimulation. Cortisol plasma levels in both groups were higher at birth (P<0.05) compared to the later sampling times. No differences were observed between the two groups indicating that the induction protocol used does not seem to result in premature foals.

Morphological features of the placenta at term in the Martina Franca donkey.

This study was designed to establish the morphological features of the placenta of the Martina Franca jenny. Ten placentas were harvested at the time of foal delivery and examined both for gross and histological characteristics. The following factors were determined: the total weight and volume of the placenta and its components, the surface area of the allantochorion, umbilical cord length and site of insertion, and the diameter of the umbilical cord vessels and vascular pattern type. The weight of the placenta was similar to previously reported for ponies, and represented 12% of foal birth weight. Umbilical cord length was similar to that in the horse but longer than in the pony, while cord weight was intermediate between the two. In a histological examination, numerous strong villi were observed at sites corresponding to the non-pregnant and pregnant horn and uterine body. No villi were detected in the area overlying the cervical star. Despite obvious similarities between the donkey and horse placenta, specific morphological features do exist, and are possibly related to the differences in length of gestation.

Luteal function in jennies following PGF(2alpha) treatment 3 days after ovulation.

Native PGF(2alpha) and its analogs have been used in the horse mare to manipulate ovarian activity, primarily as luteolytic agents to induce estrus. Despite numerous studies on the effects of these luteolysins in the mare, to date only a single investigation has been conducted in the jenny. The aim of this study was to evaluate the response of the corpus luteum (CL) to a single dose of PGF(2alpha) given 3 days (72h) after ovulation and to establish the plasma progesterone (P4) profile from pre-treatment to post-treatment ovulation in the Martina Franca donkey. Twenty-two jennies were ultrasonographically monitored and treated 72h after the detection of ovulation with 0.075 mg i.m. of R-cloprostenol. From the day of ovulation until ovulation post-treatment, blood was collected daily for P4 determination by enhanced luminescence immunoassay. All the jennies except one, exhibited behavioral signs of PGF(2alpha)-induced estrus within 4 days of treatment lasting 5.4+/-1.16 days. Post-treatment ovulation was also hastened, reducing the interovulatory interval (9.6 days). In response to treatment, plasma P4 concentrations fell to estrus levels and then remained constant until the next ovulation in all but the non-responding animal. Our findings indicate that PGF(2alpha) treatment on Day 3 post-ovulation causes the functional regression of the CL in the jenny, reflected both by the rapid induction of estrus and ovulation and by an abrupt drop in circulating P4 concentrations.

Electrolytes changes in mammary secretions before foaling in jennies.

No knowledge regarding the peripartum changes in mammary secretions in the jenny are presently available in literature. In the mare, instead, several studies report the role of these changes as indicators of foetal readiness for birth and impending parturition. This experiment was designed to determine calcium, sodium, potassium concentrations, and the value of sodium/potassium ratio in mammary fluids during prepartum in the jenny. Samples were daily collected by hand milking, after mammary gland size increased noticeably, from 17 Martina Franca jennies. Prepartum mammary secretions were analysed every other day between day 10 and day 2 antepartum, and then once a day from the day before to the day of parturition. Calcium concentration showed a significant increase between day 10 and day 6 antepartum and then between day 6 and days 4 and 2. Afterwards, another statistical significant increase was observed at parturition. Sodium concentration significantly decreased from day 10 to day 2 prepartum. Potassium concentration significantly increased between day 10 and day 8 before parturition, then showed a further increase at day 4, followed by none significant changes until foaling. All jennies showed a reversal in sodium/potassium ratio between 2 days antepartum and the day before. In conclusion, the evaluation of mammary fluid calcium concentrations and the reversal of sodium/potassium ratio could be used as good indicators of foetal maturity in the jenny. As far as the prediction of parturition is concerned, the reversal of sodium/potassium ratio is the best parameter, since it was detected 48-24 h before parturition in all considered animals.

Efficacy of hCG and GnRH for inducing ovulation in the jenny.

Knowledge about management of ovulation in the donkey is limited compared to that in the horse. This experiment was designed to evaluate the efficacy of injecting single doses of lecirelin (a GnRH-analogue) or of hCG to induce ovulation in the jenny and to determine whether effects are dependent upon follicular diameter at time of injection. Ovarian activity and follicular growth were monitored by rectal ultrasonography. Jennies were randomly allotted to the following groups: Group GnRH, treated with 100 microg lecirelin; Group hCG, treated with 2500 IU hCG; Group C, untreated and monitored for spontaneous ovulation. Animals were also categorized into subgroups depending upon follicular diameter: 30-35 mm (GnRH-1, hCG-1 and C-1) or 36-40 mm (GnRH-2, hCG-2 and C-2). Jennies in the two hormone treatment groups did not differ significantly for time from treatment to ovulation, but there was a significant reduction in time to ovulation as follicle size at treatment increased. Jennies treated with either lecirelin or hCG had significantly smaller follicle size at ovulation than jennies in the Control groups that underwent spontaneous ovulation. Treatment groups did not differ significantly in the proportion of jennies that ovulated within 48 h of injection or between 25 and 48 h following injection. These results highlight the usefulness of lecirelin for induction and synchronization of ovulation in the jenny, particularly since it would avoid the risk of reduced hCG response in reproductive management programs in which that hormone was repeatedly used.

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.

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.

Insulin-like growth factor I: could it be a marker of prematurity in the foal?

Insulin-like growth factor (IGF)-I represents one of the most important growth regulators, playing a central role in fetal and neonatal growth. Plasma IGF-I levels increase rapidly after birth, and they are influenced by numerous factors, including sex, age, nutritional state, and premature birth. The aims of this study were: (1) to evaluate the IGF-I plasma profile in healthy newborn foals during the first 2 weeks of life; (2) to assess the possible influence of sex and birth weight on this hormone; (3) to analyze the percentage increment of IGF-I values in healthy foals; (4) to evaluate the influence of prematurity on IGF-I profile; (5) to verify the role of IGF-I as a diagnostic marker of prematurity; and (6) to analyze the percentage increment of IGF-I in premature foals. Thirty-four healthy term foals were enrolled as the control group and from each foal plasma was collected within 6 hours from birth, at 12 hours, daily from Day 1 to Day 7, and at Days 10 and 14 after birth. Eleven foals aged younger than 1 week and diagnosed as premature and hospitalized at a Equine Perinatology Unit were also enrolled; from each foal plasma was collected daily from the day of admission to discharge or death. Insulin-like growth factor I was analyzed by RIA. In the control group, an increasing trend of IGF-I concentrations was found, with higher values from Day 4 to 10 compared with data obtained at less than 6 hours of life, and from Day 5 to 10 compared with 12 and 24 hours and 3 days. No differences were found in healthy foals analyzed in relation to birth weight and sex. In premature foals an increasing trend was observed but no statistical differences were found among sampling times, and no differences were found between healthy and premature foals. The IGF value in premature foals at admission was always higher compared with the lowest recorded level in healthy age-matched foals, thus this parameter does not seem to have a diagnostic role for prematurity in foals. Finally, the evaluation of the percentage increment of IGF-I concentrations showed a significant increase in full-term foals on Day 5, 6, 7, and 10 compared with 12 and 24 hours, and no differences were observed in premature foals. In conclusion, prematurity in newborn foals seems to affect only partially IGF-I plasma concentrations and it does not seem to be a reliable marker for this pathological condition.

Influence of season on testicular morphometry and semen characteristics in Martina Franca jackasses.

As other European donkey breeds, Martina Franca could be considered an endangered breed because of the population number (48 jackasses and 515 jennies in 2011). To increase donkey population and breed biodiversity preservation, several research projects on donkey reproduction have been performed; however reproductive seasonality has been only partially investigated in jackasses. For this reason the aim of this study was to investigate the effect of seasons and of long (spring to summer; SS) and short (autumn to winter; AW) day length periods, on reproductive physiology of Martina Franca jackasses, in particular on: (1) testicular morphometric characteristics, (2) behavior, through the evaluation of the reaction time, and finally, (3) semen characteristics. Seven adult and reproductively mature Martina Franca jackasses were enrolled. For each jackass, a morphometric evaluation of both testes was performed once for every season, before the first seasonal collection. Semen was collected, by artificial vagina, weekly for a whole year (52 collections per donkey); at each collection the reaction time was recorded and a complete semen evaluation was performed immediately after collection. No differences in testicular measures were observed neither between left and right testis nor during the four seasons. A lower reaction time was observed in spring and summer compared with autumn and winter and during the period SS compared with AW. Total volume was significantly higher in winter compared with all the other seasons; gel-free volume was higher in winter compared with summer and autumn. Mean sperm concentration was significantly lower in winter compared with spring and summer and in AW compared with SS. Total and progressive motility and membrane integrity did not show any significant difference between season and between SS and AW. A lower average path velocity was observed in autumn compared with spring and summer, and in summer a higher straight line velocity compared with spring and autumn, and a lower curvilinear velocity compared with winter were found. No differences in amplitude of lateral head displacement, beat cross frequency, straightness and linearity were observed neither between seasons nor in SS versus AW. Seminal pH did not show any seasonal difference; also sperm morphology did not show any significant difference during the whole year. This study demonstrated that under our climatic and management conditions Martina Franca donkey stallions do not show significant differences in testicular morphometric characteristics during the year, even if a certain seasonal influence exists on reaction time and some seminal parameters. Considering the variable temperature and photoperiod during the four seasons where the study was performed, this lack of substantial differences could be related to a characteristic of the Martina Franca breed.

How type of parturition and health status influence hormonal and metabolic profiles in newborn foals.

Thyroid hormones, insulin growth factor I (IGF-I) and non-esterified fatty acids (NEFA) represent important hormonal and metabolic factors associated with perinatal growth and maturation. Their action could be influenced by the type of parturition and the health status of the foal and therefore the aim of this work is to evaluate their plasma concentrations in newborn foals during the first 2 wks of life. Three groups of subjects were enrolled: 15 healthy foals born by spontaneous parturition, 24 healthy foals born by induced parturition and 26 pathologic foals. From each of the healthy foals, blood was collected at 10, 20 and 30 minutes, 3 and 12 hours from birth, daily from Day 1 to Day 7, and at Day 10 and 14 of life. In pathologic foals samples were collected twice a day from the day of admission at the hospital until the day of discharge or death. Thyroid hormones (T3 and T4) and IGF-I were analyzed by radioimmunoassay and NEFA by enzymatic-colorimetric methods. In all the three groups a declining trend of T3 and T4 plasma concentrations was detectable, with lower levels in the pathologic group compared to healthy foals. Spontaneous foals showed higher levels of T3 at 7 d compared to induced foals, while T4 levels were higher in spontaneous vs. induced foals before 6 h of life, at three and seven days. IGF-I showed increasing plasma concentrations in all three considered groups. No differences were found between healthy and pathologic foals. NEFA in spontaneous and induced healthy foals showed a declining trend with higher levels during the first hours of life. Pathologic foals presented higher levels compared to spontaneous foals only at 24 h and 10 d. These data suggest that the type of foaling could influence the reference ranges for thyroid hormones. Moreover, pathologic foals showed some hormonal and metabolic differences related to their health status. Above all changes of thyroid hormones levels, early in postnatal life, could be a cause, and not only a consequence, of the diseased condition of these foals.

PGF(2α), LH, testosterone, oestrone sulphate, and cortisol plasma concentrations around sexual stimulation in jackass.

Many hormones are involved in the regulation of male reproductive functions, controlling sexual behavior, and influencing sexual arousal, the onset of erection and ejaculation, and the post-ejaculatory detumescence. The aims of this study were to analyze the plasma concentrations of 15-ketodihydro-PGF(2α) (PGFM), LH, testosterone (T), oestrone sulphate (OS), and cortisol (C) in relation to sexual stimulation and to evaluate the possible correlations among circulating hormones and between hormones and semen characteristics in the donkey stallion. Thirteen sexually experienced Martina Franca jackass of proven fertility were enrolled and semen was collected through an artificial vagina. Plasma samples were collected at 12, 9, 6 and 3 min before oestrous jenny exposure, at the first erection in the mating arena in the presence of an oestrous jenny, during ejaculation, at dismounting, 3, 6, 9 and 12 min after ejaculation in box, and then every 10 min during the following 50 min. PGFM showed an increasing trend with significant differences between the pre-ejaculatory and post-ejaculatory period, suggesting a role of this hormone in the control of ejaculation. LH showed a significantly higher concentration at ejaculation compared to last samples, while T showed significantly higher levels at erection, ejaculation and dismounting, probably for its influence on these processes and on sexual behavior. Finally, OS did not show any difference in the period of observation, while C presented a significant increase only 22 minutes after erection. The only hormonal correlation found was a positive one between LH and T at erection and dismounting, while T and OS were positively correlated with total and progressive motility, respectively.

An Apgar scoring system for routine assessment of newborn puppy viability and short-term survival prognosis.

The Apgar scoring system is an easy and reliable method for evaluating both human and animal neonates. However, its use is not widespread in veterinary medicine. The current study assessed a modified Apgar scoring system for routine evaluation of newborn puppies. Heart rate, respiratory effort, reflex irritability, motility, and mucus color have been evaluated in the score. Specifically, we used 5 min after birth Apgar score to assess newborn viability and short-term survival prognosis, as well as related characteristics, in 193 puppies from 42 litters, 65 born by spontaneous delivery, 66 by assisted delivery, and 62 by cesarean section. The percentage of puppies that were dead 2h after birth was higher in the 4 to 6 Apgar score group versus that in the 7 to 10 score group (P<0.01) and in the 0 to 3 score group versus that in the 7 to 10 score group (P<0.0001). Delivery method did not affect survival. There was a marked reduction in the number of puppies searching for the mammary gland in the 0 to 3 and 4 to 6 Apgar score groups compared with that in the 7 to 10 score group (P<0.0001); there was a difference between the 0 to 3 and the 4 to 6 score groups as well (P<0.05). Suckling/swallowing reflexes were present in fewer puppies in the 0 to 3 and 4 to 6 score groups compared with that in the 7 to 10 group (P<0.0001), with no significant differences between the 0 to 3 and the 4 to 6 score groups.

Concentrations of 15-ketodihydro-PGF2alpha, cortisol, and progesterone in the plasma of healthy and pathologic newborn foals.

Information regarding the plasma hormone profiles of prostaglandins (PGs), cortisol (C), and progesterone (P4) during pathologic processes in newborn foals is scarce. The aim of this study was to determine the plasma concentrations of these hormones in diseased foals (n=40) and healthy at-term foals (n=24) (Equus caballus) during the first 2 weeks of life. Blood samples were collected daily, before any treatment with nonsteroidal drugs in diseased foals, and plasma was analyzed by radioimmunoassay. 15-Ketodihydro-PGF(2alpha) (PGM) was consistently higher in diseased foals than in healthy foals, probably related to roles of PGs in completing organ maturation and/or the presence of oxidative stress or inflammation. Similar trends were observed for C and P4. In diseased newborns, only PGM was significantly higher in nonsurviving foals, although C showed a similar profile. When specific diseases were considered, the levels of PGM and C were lower in premature foals at 12h of life, whereas the concentration of P4 was higher than in controls. The results of this study demonstrate the differences in plasma hormone levels between healthy and pathologic newborn foals, particularly during the first 2 d of life, probably reflecting the inability of diseased foals to cope with the transition between fetal and neonatal life.