Ozone therapy in veterinary medicine: A review.

Ozone (O3) is a triatomic form of oxygen. As O3 rapidly dissociates into water and releases a reactive form of oxygen that may oxidize cells, the gas mixture of O3/O2 is used in medicine. ATP is widely available for cellular activity. O3 can be administered via the systemic and local routes. Although O3 is known as one of the most powerful oxidants, it also promotes antioxidant enzymes. Additionally, it stimulates some of the cells of the immune system and inactivates pathogens, including bacteria, fungi, yeasts, protozoa, and viruses. Owing to these activities, O3 is used to improve several diseases, both in human and in veterinary medicine. Considering the wide scope of O3 application, the aim of this review was to reiterate the mechanisms of action of O3 and its utilization in different mammalian species (bovine, ovine-caprine, equine, canine, porcine).

Oxidative state in the estrus cycle of the buffaloes: a preliminary study.

The oxidative status, the relationship between oxidizing agents (free radicals) and antioxidant system, is able to influence the correct performance of the estrus cycle. The aim of the experimentation was to define a first range of physiological values of reactive oxygen species (ROS) and the biological antioxidant potential (BAP), found during the phases of the buffalo estrus cycle. In this study, blood samples were taken from 30 buffaloes in various phases of the estrus cycle (estrus, diestrus, anestrus), on which serum determinations, by a photometer, of BAP and ROS have been carried out. The highest ROS values were detected during estrus and this is due to the primary role they play in determining ovulation. The highest levels of BAP have been found in diestrus, when they perform protective action against oxidative damage in the ovaries and uterus.

Alpha-fetoprotein in animal reproduction.

Alpha-fetoprotein (AFP) is a serum glycoprotein with structural and physico-chemical properties similar to albumin. However, the exact physiological functions of AFP remain unknown; those known to date include markers to pathological conditions including neoplastic and non-neoplastic diseases, antioxidant effects, growth regulator in different cells and in cancer, immune response modulator, and carrier for fatty acids and oestrogens. This review aimed to present an overview of the different functions of AFP, particularly its role in the sexual differentiation of the hypothalamus, because its ability to bind oestrogens prevents their passage to the brain, where they inhibit the surge centre development. AFP and anti-Mullerian hormone are known to be involved in the development of freemartins, or genetically female foetuses masculinised in the presence of a male co-twin.

Role of homocysteine metabolism in animal reproduction: A review.

Homocysteine (Hcy) is a thiol-containing essential amino acid, important for the growth of cells and tissues. Several hypotheses exist regarding Hcy toxicity in humans; Hcy is involved in protein structural modifications, oxidative stress, and neurotoxicity induction and is therefore associated with several pathological conditions in humans. In veterinary science, knowledge regarding Hcy has increased recently due to several studies; however, many aspects remain undiscovered. Many details remain unknown regarding the effect of Hcy levels on pregnancy and the optimal management of pathological conditions associated with Hcy levels during pregnancy in various species. In this review, we aimed to compile various studies on Hcy metabolism to elucidate its current status in the veterinary field, particularly for ovine, bovine, equine, porcine, canine, and feline species.

Scopolamine for uterine involution of dairy cows.

The objective of this study was to evaluate the effect of administration of scopolamine (a parasympatholytic drug) immediately after delivery, on uterine involution in dairy cows. The cows were divided into two groups: group T (treated with scopolamine butylbromide 40 mg/100 kg P.V., within 24 h of delivery) and group C (treated with 2 ml/q of saline solution). The cows were monitored at T0, within 24 h of delivery, T7, T14, T28 and T40 respectively 7th, 14th, 28th and 40th day postpartum, by measuring the levels of hydroxyproline (HYP), an important marker of uterine involution; by ultrasound examination for the measurement of the diameter of uterine horns and of blood flow in the middle uterine artery; and by evaluation of reproductive indices. HYP showed higher concentrations in the T group than in the C group at all times, but the difference was significant at T7 and T40. Uterine diameters were reduced from T7 to T40, with lower values in the T group, as compared to the C group; the pulsatility index (PI) of the middle uterine artery increased in the T group compared to the C group, with a statistically significant difference at T7. There was a better conception-delivery interval and higher pregnancy rate in the T group compared to the C group. It is likely that scopolamine blocked the uterine contractions in postpartum, for the duration of its half-life. After cessation of its pharmacological effect, the uterus may have started to contract more effectively. These results suggested that scopolamine could be useful as a pharmacological approach to postpartum management.

Contractile effect of rifaximin on bovine uterus in the presence of steroid hormone antagonists.

This in vitro study investigated the modulatory effect of rifaximin on bovine uterus contractility, in both phases of the oestrous cycle, with and without the steroid hormones that are predominant in the respective phases: oestrogen in the follicular phase and progesterone in the luteal phase. The procedures were conducted in an isolated organ bath by using rifaximin alone (10-4 M) and in association with the steroid hormone antagonists (10-5 M) tamoxifen (oestrogen antagonist) in the follicular phase and mifepristone (progesterone antagonist) in the luteal phase. The results indicated that rifaximin can stimulate uterine contractility. Indeed, the administration of rifaximin in the presence of tamoxifen or mifepristone increased the tonic activity of the uterus in both phases of the cycle. This result is clinically significant because rifaximin might also enable, in vivo, a simultaneous increase in uterine cleaning and the antimicrobial action against bacteria during the first 14 days postpartum, during the development of acute metritis.

Scopolamine butylbromide decreases the xylazine-mediated contractility in bovine pregnant uteri.

The objective of this in vitro study was to evaluate and compare the effects of scopolamine butylbromide (Spasmolax®) on xylazine-sensitized bovine pregnant uterine strips, at different stages of pregnancy. The procedures were carried out in isolated organ bath. Uterine motility, expressed with amplitude, frequency of contractions as well as the area under the curve, was recorded in different stages of pregnancy and data were collected at 5-min intervals starting 5 min before treatment until 10 min after treatment (5-min after xylazine administration and 5-min after scopolamine butylbromide addition). The results suggest that scopolamine butylbromide might decrease the tonic effect induced by xylazine on bovine pregnant uteri from 0-30 days to 240-270 days.

Effects of d-cloprostenol on different layers and regions of the bovine uterus during the follicular and luteal phases.

INTRODUCTION: D-cloprostenol is a synthetic PGF2α, commonly used in bovine reproduction, that increases myometral contractility. However, little is known regarding its contractile behaviour and how it depends on the reproductive state and uterine topography (regions and muscular layers). HYPOTHESIS AND OBJECTIVES: These aspects would affect the action of d-cloprostenol on the uterus. Therefore, we hypothesize a possible use of this drug at the time of artificial insemination, to improve conception rates and, in the post-partum, in order to accelerate uterine involution in dairy cattle. The purpose of the present study was to investigates the modulatory effect of d-cloprostenol on contractility of the bovine uterine region (horn and corpus) and their muscle layers (circular and longitudinal), in follicular and luteal phases. To our knowledge, motility effects of d-cloprostenol on different regions from healthy bovine uterus have not been described up to now. MATERIALS AND METHODS: Uterine specimens were collected from uterine body and horn of cattle in the follicular (n = 28) and luteal phase (n = 32) of the estrous cycle at slaughter. Two strips were prepared from each regions corresponding to the circular and the longitudinal muscular layers, respectively. Samples were cultured in an organ bath, exposed to synthetic prostaglandin (1 µM d-cloprostenol) and their contractile activity was monitored for 10 min. The functionality of the strip throughout the experiment was tested by a dose of carbachol (10-5 M). RESULTS: The mean basal amplitude of contractions was higher in the follicular compared to the luteal phase in uterine horn samples, but not in muscles collected from the uterine body. The amplitude of contractions increased after d-cloprostenol administration in all tissues with a greater increase in samples from cattle in the follicular phase. The frequency of contractions increased after d-cloprostenol administration in longitudinal but not in circular fibrees. CONCLUSION: The contractile responses to d-cloprostenol in both horn and corpus were strongest in the circular muscles but weak in the longitudinal muscles.

Effect of neostigmine on contractility of equine pre-ovulatory follicles: An in vitro study.

In this study, the Authors investigated the modulatory effect of three single doses (10-6, 10-5, and 10-4 M) of neostigmine on the spontaneous contractility of equine pre-ovulatory follicles in an isolated organ bath, to establish the relationship between this acetylcholinesterase inhibitor and ovulation, in the mare. The results indicate that neostigmine increases pre-ovulatory contractility in equine follicles at each dose, but in a different manner. Indeed, the rise in contractility induced by neostigmine at 10-6 M and 10-4 M was phasic, while at 10-5 M it was tonic. The data obtained indicate possible implications of these drugs in the pharmacological modulation of equine ovulation.

Dynamics of the progesterone and cholesterol concentrations within the bovine corpus luteum cavity.

The aim of this study was to examine the concentrations of progesterone (P4) and cholesterol (CHOL) in fluid of cavitary corpus luteum (CL) and in serum of dairy cattle. Cavitary corpora lutea, collected from cows at a local abattoir, were divided in four stages of development, based on days of the estrous cycle (stage I: 1-4days; stage II: 5-10days; stage III: 11-17days; stage IV: 18-20). Fluid of the their cavity was aspirated and P4 and CHOL concentrations were evaluated. The concentrations of P4 in CL cavity fluids were very high (1640-4666.67ng/mL) and showed a peak at the stage III. CHOL values were similar to those of serum and the highest levels were found in the stage I.

Application of an innovative technique for unilateral ovariectomy in dairy cows.

This report describes an innovative unilateral ovariectomy technique for the surgical treatment of follicular cysts in dairy cows. Five cows, with a diagnosis of follicular cysts, were submitted to unilateral ovariectomy via a paralumbar fossa approach, in standing position. The linear cutter proved useful for clamping the ovary, stapling the vessels and cutting in one stroke, thus reducing surgery times. After unilateral ovariectomy, all cows underwent heat 7-30 days after surgery, and three cows became pregnant. All cows showed increased milk production. The results of this case report showed that flank ovariectomy, performed in this way, is an efficient, low-risk technique for surgical treatment of follicular cysts in dairy cows.

Lidocaine decreases the xylazine-evoked contractility in pregnant cows.

The objective of this in vitro study was to evaluate and compare the effects of xylazine on basal uterine contractility of bovine pregnant uterine strips and that of lidocaine on xylazine-sensitized bovine pregnant uterine strips, at different stages of pregnancy. Basal contractility was evaluated in an isolated organ bath and the functionality of the strips throughout the experiment was evaluated using a dose of carbachol (10(-5)M). Uterine motility, expressed with amplitude, frequency of contractions as well as the area under the curve, was recorded in different stages of pregnancy and data were collected at 15-min intervals (5-min before and 5-min after xylazine administration and 5-min after lidocaine addition on the plateau contraction induced by xylazine). Uterine motility increased in all the stages of pregnancy after xylazine addition and gradually decreased after treatment with lidocaine. These data suggest that lidocaine might decrease the tonic effect induced by xylazine on bovine pregnant uteri.

Oxytocin plus antibiotics: A synergism of potentiation to enhance bovine uterine contractility.

This in vitro study investigates the modulatory effect of three antibiotics (amoxicillin, enrofloxacin, and rifaximin) on contractility of the bovine uterine tissue, in follicular and luteal phases. The evaluation of the effects of these antibiotics (10(-4) M) was performed on oxytocin-induced contractility. The decision to test these antibiotics with the oxytocin (10(-6) M) comes from the reported ability of these combinations of hinder the antibiotic resistance and the formation of bacterial biofilms. The procedures were carried out in isolated organ bath, and the contractile functionality of the strip throughout the experiment was evaluated after a dose of carbachol (10(-5) M). The results demonstrate the different modulatory activity of these antibiotics, on the plateau of contraction induced by oxytocin, in both phases of the estrus cycle. The differing individual antibiotic effects of our testing made it possible to identify, only in some cases. Rifaximin in the follicular phase and enrofloxacin in both phases of the estrous cycle, induced a synergistic enhancement (potentiation) of uterine strip contraction induced by oxytocin. This result is thought important because these associations might enable, in vivo, a simultaneous increase of uterine cleaning and the antimicrobial action on bacteria in planktonic form and of those organized in biofilms.

Effects of kisspeptin-10 on in vitro proliferation and kisspeptin receptor expression in primary epithelial cell cultures isolated from bovine placental cotyledons of fetuses at the first trimester of pregnancy.

Kisspeptin (Kp) and Kiss-1 receptor (Kiss-1R) expressions have been reported to be in the placenta, and a possible involvement of the Kiss-1R/Kps system in regulating trophoblast invasion and proliferation has been hypothesized. The aim of the present study was to investigate whether Kiss-1R activation by kisspeptin-10 (Kp-10) could modulate in vitro proliferation and progesterone (P4) secretion of bovine primary placental cell lines isolated from cotyledons of fetuses in the first trimester of pregnancy. The involvement of Kiss-1R in the cell responses observed was also analyzed. Uteri from cows at the first trimester of pregnancy were obtained from local abattoirs. Fetal cotyledon fragments were digested with collagenase in low glucose Dulbecco's Modified Eagle's Medium and cell lines were isolated. After being characterized for epithelial polygonal morphology, the presence of binucleate cells, male gender, and the expression of cytokeratin and zona occludens 2, cell lines were cultured in a low glucose Dulbecco's Modified Eagle's Medium-based expansion medium in the presence of 0.01, 0.1, 1, and 10 µM Kp-10. Control cells were cultured in the absence of Kp-10. Cell population doubling time was evaluated for each culture passage (P) from P1 to P10. Cells were tested for Kiss-1R mRNA expression analysis by real-time reverse transcription-polymerase chain reaction, and culture media were analyzed for P4 concentration by radioimmunoassay. Kisspeptin-10 modulated in vitro proliferation of epithelial cell lines isolated from cotyledons recovered from bovine fetuses in the first trimester of pregnancy. Inhibitory (line A) or stimulatory (line B) effects of Kp-10 on cell proliferation were found in different cell lines and observed cell responses were found to be related to Kiss-1R mRNA levels. Inhibition of cell proliferation matched with not significant variation of Kiss-1R expression, whereas stimulation of cell proliferation was found to be related to Kiss-1R upregulation. In both cell lines, no effect of Kp-10 on P4 secretion was found at any tested concentration. These results lead to the conclusion that the Kiss-1R/Kps system is involved in the regulation of cell proliferation of bovine placental cotyledon cell lines isolated at the first trimester of pregnancy but, at this gestational stage, it may not be involved in modulating placental P4 secretion.

Modulatory effect of three antibiotics on uterus bovine contractility in vitro and likely therapeutic approaches in reproduction.

This in vitro study investigates the modulatory effect of three antibiotics (amoxicillin, enrofloxacin, and rifaximin) on contractility of the bovine uterine tissue in follicular and luteal phases. The effects of these antibiotics at three single doses (10(-6), 10(-5), and 10(-4) M) on their basal contractility were evaluated in isolated organ bath. The functionality of the strip throughout the experiment was evaluated by a dose of carbachol (10(-5) M); the obtained effect had to be repeatable (difference of ≤20%) that is comparable to that induced by the previous administration of the same substance. The results demonstrate the different modulatory activities of these antibiotics on uterine contractility in follicular and luteal phases. The effects induced by amoxicillin and enrofloxacin are opposite: the first relaxes and the second increases the uterine contractility in both cycle phases. Instead, the activity of rifaximin varies depending on the phase of estrous cycle: it increases in the follicular phase and relaxes in the luteal phase. The obtained data provide the hypothesis of possible implications of these drugs in the pharmacologic modulation of uterine contractions. Their action at this level, associated with their specific antimicrobial effects, could suggest using these antibiotics for the treatment of diseases related to postpartum or infections that may occur in pregnant cattle, by virtue of their effects on myometrial contractility too.

In vitro effect of isotocin on ovarian tunica albuginea contractility of gilthead seabream (Sparus aurata L.) in different reproductive conditions.

Contractions of ovarian tunica albuginea, the teleostean cystovary wall layer containing smooth muscle fibres, facilitate oocytes and fluids movements within the ovary, oocytes ovulation and spawning. Fish isotocin, the homologue hormone of mammalian oxytocin, plays a significant role in ovulation, oviduct contraction and spawning. In the present study, ovarian wall spontaneous contraction, as well as isotocin in vitro effect on tunica albuginea contractility, was analysed in female seabream in different reproductive conditions: vitellogenesis, regressing (post-spawning) and extensive atresia. Tunica albuginea spontaneous contractility was recorded using ovary wall strips mounted in an organ bath containing modified Ringer's solution. The strips were then exposed to cumulative doses of isotocin (6, 30, 60 µg/ml). Female seabream in regressing condition exhibited the highest level of tunica albuginea spontaneous contraction amplitude compared with the other two groups. Only fish in vitellogenesis state showed a significant increase in contraction amplitude after isotocin administration at the dose of 30 µg/ml. The same group exhibited also a significant isotocin dose-dependent decrease in the contractile frequency. These results confirm the involvement of isotocin in stimulating tunica albuginea contractile activity during the oestrogen-regulated phase of vitellogenesis, whereas the absence of significant effects of isotocin on ovarian contractility in fish at the regressing state might be ascribed to the occurrence of a contractile activity autonomously regulated by the internal pacemaker system. The absence of exposed isotocin receptors could explain the lack of effects of the isotocin administration in seabream showed extensive atresia of the follicular cells.

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.

Oxidative stress in neonatology: a review.

Free radicals are highly reactive oxidizing agents containing one or more unpaired electrons. Both in human and veterinary neonathology, it is generally accepted that oxidative stress functions as an important catalysator of neonatal disease. Soon after birth, many sudden physiological and environmental conditions make the newborn vulnerable for the negative effects of oxidative stress, which potentially can impair neonatal vitality. As a clinician, it is important to have in depth knowledge about factors affecting maternal/neonatal oxidative status and the cascades of events that enrol when the neonate is subjected to oxidative stress. This report aims at providing clinicians with an up-to-date review about oxidative stress in neonates across animal species. It will be emphasized which handlings and treatments that are applied during neonatal care or resuscitation can actually impose oxidative stress upon the neonate. Views and opinions about maternal and/or neonatal antioxydative therapy will be shared.

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.