Ovarian contractility in reared gilthead seabream (Sparus aurata L.) in different phases of the reproductive cycle.

Spontaneous ovarian tunica albuginea contractility was evaluated in gilthead seabream (Sparus aurata L.) at different phases of the reproductive cycle. Fourteen adult females were sampled from February to November 2012 in a commercial fish farm, and ovaries were removed and processed for histological and contractility analyses. Fish reproductive stages were evaluated on haematoxylin-eosin-stained ovary sections or by simple macroscopic observation of hydrated oocytes in spawning individuals. Tunica albuginea spontaneous contractility was recorded by using ovary wall strips mounted in an organ bath containing modified Ringer's solution. Ovary macro- and microscopic analyses allowed the identification of three different reproductive conditions: vitellogenesis, spawning and regressing. The gilthead seabream tunica albuginea was capable to contract spontaneously, and significant differences were found in mean contraction amplitude among the three reproductive states, with the highest value recorded in individuals in regressing condition and the lowest in individuals at spawning stage. No differences in mean contractility frequency among the three different groups were found. Possible involvement of spontaneous contractility in facilitating developing follicle advancement towards the ovarian lumen within the ovary and in supporting recovery of regressing ovaries may be hypothesized. The low contractility observed during the final oocyte maturation and spawning phases does not seem to support a role of tunica albuginea during ovulation, which could conversely involve theca cell contraction. Alternatively, possible single instantaneous contractions of tunica albuginea muscle fibres, not detected in the present study, could occur during ovulation in response to neuro-hormonal stimulations; a role of abdominal wall musculature in ovary "squeezing" and consequent release of ovulated eggs cannot be excluded.

Blood and milk oxidative status after administration of different antioxidants during early postpartum in dairy cows.

This paper describes serum and milk oxidative profiles during early postpartum (10th-16th day) of dairy cows, after three different treatments: ß-carotene and α-tocopherol (Dalmavital®, Fatro, Italy) (VAE Group); L-carnitine, d, L-acetylmethionine, cyanocobalamin (Adiuxan®, Ceva Vetem, Italy) (LCAR Group) and Dalmavital®+Adiuxan® (VAE/LCAR Group). Antioxidant administrations resulted in a general reduction in ROS concentrations which decreased more after a single drug administration (VAE and LCAR Groups) vitamin A and E serum levels increased in the three treated groups compared to control one (p < 0.05). Milk lipoperoxide content was statistically lower in treated cows than in control ones, whereas, as to vitamins, only vitamin E increased in all the treated groups compared to control. Taken together, the results of this study confirm the efficacy of antioxidant administration in the early postpartum of dairy cows and suggest that an excess in antioxidant administration may result, paradoxically, in an increase in ROS generation.

The expression of leptin receptor in the ovary of the queen: leptin receptor expression in queen ovary.

Leptin is a Ob gene product secreted mainly by adipose tissue. Several reports showed leptin production by other tissue including the ovary. The action of leptin is mediated upon binding to its receptor widely expressed in reproductive tissues in different species. In fact, there are growing evidences that leptin plays an important role in the modulation of reproductive functions. Therefore, the aim of this study was to evaluate in the queen, the expression of leptin receptor during the functional ovarian cycle and pregnancy. We found that the ovaries of the queen express leptin receptor in all the examined phases. The highest leptin receptor expression was found in the luteal phase (pseudopregnancy, pregnancy) compared to other phases of the cycle (anestrus, proestrus, estrus). The variations in the expression of leptin receptor suggest a likely implication of leptin in the modulation of ovarian activity, in the examined species.

Leptin and queen ovary: new insights about ovulation.

Leptin has been proven to be crucial in the ovulatory process. Aims of this study are to assess the expression of leptin receptor (Ob-R) in the ovaries of queens at estrus and to evaluate the capability of leptin in modulating ovarian contractility in vitro. Right ovaries underwent immunoblot analysis. Left ovaries were mounted in an organ bath under physiological condition and exposed to murine leptin (10(-6) M). Immunoblot analysis showed that the queen ovary expresses leptin receptor at estrus. Leptin at the dose of 10(-6) M significantly reduced the contractile activity of the ovary. The presence of ovarian Ob-R and leptin inhibitory effects on ovarian contractility suggest leptin implication in the modulation of ovarian activity, as well as in ovulatory disorders.

Renin and ovarian vascularization in cows with follicular cysts after epidural administration of a GnRH analogue.

The ovarian renin-angiotensin system may play an important role in follicular growth and maturation, as well as in the process of ovulation. The aim of this study was to investigate the effects of administration of a GnRH analogue to cows with ovarian follicular cysts on plasma renin concentrations and ovarian vascularization. This study was performed with 60 Friesian cows, which were diagnosed with follicular cysts, and randomly allocated into two groups: group A (treatment; n=30) received 2ml of lecirelin (Dalmarelin((R)) - Fatro), per head via sacro-coccygeal epidural, and group B (control; n=30) received 2ml saline solution (0.9% NaCl) per head by the same route. Blood samples were immediately collected prior to administration (T0) and then 24h (T1), 48h (T2) and 8 days (T3) after administration of the treatment, for both groups. Ovarian vascularization was evaluated utilizing Power Doppler on these same days in 10 animals from each group. The number of pixels detected by Power Doppler was used as an indicator of the degree of vascularization. Plasma renin concentrations remained relatively constant for the control (group B) animals, but increased as the sampling period progressed (NS) for the treated cows (group A). Similarly, there were no changes in ovarian vascularization (number of pixels) for the control cows, but vascularization increased throughout the sampling period in the treated animals. The number of pixels associated with cysts was significantly higher for treated compared to control cows at 24h after treatment (P<0.001). The epidural administration of a GnRH analogue was determined to be a highly effective therapy for follicular cysts (regression occurred in 82% of treated cows within 8+/-2 days after treatment, but in none of the control cows), which also enhanced ovarian vascularization.

Influence of antibiotic therapy on serum levels of reactive oxygen species in ovariectomized bitches.

This study was conducted on 60 ovariectomized bitches. The objectives were to measure the mean reactive oxygen species (ROS) concentrations before, during and after surgery, and to investigate the effect of the administration of five different antibiotic treatments: amoxicillin, benzylpenicillin/dihydrostreptomycin, sulfametazine/sulfamerazine/sulfathiazole, enrofloxacin, lincomycin/spectinomycin. The first value recorded represented the mean ROS concentration in anestral bitches and constitutes a reference level with which to compare the subsequent measurements. After premedication, induction of anesthesia and during maintenance and surgery, ROS serum concentrations showed constant values until the end of surgery. After surgery and during antibiotic administration, an increase in ROS concentration occurred, which differed among the five groups in relation to the antibiotics employed. The lowest increases occurred in the groups treated with the combination of lincomycin/spectinomycin, and with amoxicillin; whereas the highest increases were detected in the group treated with enrofloxacin. The three other antibiotics showed an intermediate level of influence on oxidative status.

Plasma homocysteine levels in cycling, pregnant, and spayed bitches.

The aim of this study was to evaluate the physiological range of homocysteine (Hcy) exhibited by bitches during the follicular (pro-oestrus), luteal (dioestrus) and anoestrus phases of cycling animals, and in pregnant and spayed bitches. The lowest concentrations of Hcy were observed during dioestrus (3.2+/-1.27micromol/L) and in pregnant bitches (3.9+/-1.72micromol/L), and the highest concentrations during anoestrus (7.8+/-0.6micromol/L) and in spayed bitches (12.1+/-5.16micromol/L).

Intra-vesicle administration of D-cloprostenol for induction of abortion in mid-gestation bitches.

This study evaluated a new method for mid-gestation termination in the bitch, which consisted of ultrasound-guided administration of a single dose of d-cloprostenol, a PGF(2alpha) analogue, into a single gestational sac. Effects on serum progesterone concentration (P(4)) were also investigated. The study was performed between days 28 and 35 of gestation on 15 privately owned crossbred bitches, randomly divided into two groups: group A comprised 10 bitches treated with 15 microg per head d-cloprostenol diluted in 0.8 ml sterile saline (final volume 1 ml); group B comprised 5 bitches treated with 1 ml of sterile saline solution (0.9% NaCl), administered in the same way. In all bitches of group A, fetal death was successfully induced within 5 days (mean: 3.1 days, S.D. 1.2) with no clinical or behavioural complications. Mild adverse effects were observed in two bitches, each weighing less than 10 kg, including salivation, defecation and hyperventilation, which disappeared within 15 min. None of the subjects in group B aborted within 10 days post-treatment. In group A, P(4) declined 2.8 days before pregnancy termination to a mean value below 30 nmol/l (S.D. 2.9 nmol/l). However, two bitches showed a higher concentration of P(4) throughout the sampling period. Our study demonstrates that intra-vesicle administration of a single low dose of D-cloprostenol is an effective and safe technique for induction of abortion, which offers an additional option for termination of unwanted pregnancy in the mid-gestation bitch.

Concentrations of free radicals and beta-endorphins in repeat breeder cows.

Repeat breeding (RB) is one of the major problems that affect the reproductive efficiency and economy of milk production in dairy animals. So far, the etiopathogenesis of this pathology has not been defined completely. Stress has been hypothesized to be a cause of impaired reproductive efficiency. Stress may cause an overproduction of beta-endorphins and free radicals; in particular, reactive oxygen species (ROS). The aim of this work is to determine the concentrations of these substances in RB cows and to evaluate the correlation with the serum level of progesterone. The study was performed on 60 dairy cows: 26 RB and 34 control cows. Blood samples were collected on day 12 and day 16, after artificial insemination (AI) in all subjects, in order to assess the concentrations of progesterone, free radicals and beta-endorphins. The stressors, free radicals and beta-endorphins, that we considered, were higher in repeat breeders (day 12, 93.32(+/-1.91) UCarr and 0.50(+/-0.03) ng/ml; day 16, 94.42(+/-1.91) UCarr and 0.61(+/-0.03) ng/ml), with a lower level of progesterone, which probably is responsible for failure to conceive. The stress factors (free radicals and beta-endorphins) may actually enhance each other and induce an inhibition of progesterone synthesis in repeat breeders.