Intrafollicular oocyte transfer (IFOT): Potential feasibility in the ovine species.

Intra-follicular oocyte transfer (IFOT) is a promising and innovative technique for in vivo embryo production previously described for equines and bovines. The aim of this study was to assess the feasibility of IFOT in the ovine species. Two preliminary in vivo and in vitro trials were performed to test the optimal procedures and timing for IFOT. In the in vivo trial, follicular growth was monitored with transrectal ultrasonography in ten adult ewes to preliminarily determine the ovulation and ideal timing for IFOT. The in vitro trial assessed i) the optimal inner diameter of the injection needle and ii) the recovery rate and integrity of injected cumulus-oocyte complexes (COCs) after follicle aspiration. For IFOT and embryo collection, five ewes were synchronized by CIDR insertion. Forty hours after CIDR removal, in ewes under sedation and general anesthesia, the ovaries were exposed by laparotomy, and the preovulatory follicle was injected with COCs previously collected from ovaries obtained from an abattoir. At 4 h after surgery, fully recovered ewes were housed in a paddock with a ram of proven fertility. Crayon marking on ram's chest was used to detect mating. Ovulation was assessed 40 h after the transfer of oocytes by transrectal ultrasonography. On day 6 after IFOT, embryo collection was performed by uterine flushing. In the in vitro testing, injection of >5 mm follicles with a 28 G needle loaded with 30 COCs in a 5 µL volume resulted in higher recovery rates and better preservation of COCs integrity. In the in vivo trial, ultrasound scanning revealed that ovulation occurred between 60 and 72 h after CIDR removal in all animals. In one ewe subjected to IFOT, 22/24 oocytes were effectively injected into the preovulatory follicle, but no embryos were collected after flushing. In the remaining four animals, 85/102 oocytes were injected, and six cleaved embryos, 12 morulae and 1 blastocyst were collected, including native embryos. This preliminary investigation indicated that IFOT in ovine species resulted in ovulation, fimbrial capture, tubal transport of heterologous oocytes and in vivo embryo production. Further studies are needed to optimize the embryo recovery rate and develop less invasive techniques for oocyte injection and uterine flushing, such as through a laparoscopic or transcervical approach.

The Design of a New Catheter for Transcervical Artificial Insemination in Ewes.

In ovine species, transcervical artificial insemination (TCAI) is limited by the poor quality of frozen-thawed semen and by the convoluted cervical lumen hampering the passage of inseminating devices. The aim of the study was to test the efficiency of three newly designed catheters with bent tips of 3.5 mm, 5.0 mm or 8.0 mm in terms of reproductive performances (experiment 1) and to compare the results of TCAI with the best performing catheter of experiment 1 to those obtained in ewes submitted to surgical incision of cervical folds (SICF) prior to insemination (experiment 2). The following parameters were assessed: time to pass the cervix; depth of cervical penetration; site of deposition of semen; pregnancy (PR); and lambing rates (LR). The results of experiment 1 indicated that the 5.0 mm tip catheter resulted in deeper and faster TCAI and higher PR and LR compared to 3.5 mm and 8.0 mm tip catheters (p < 0.05). In experiment 2, TCAI with the 5.0 mm catheter did not differ from TCAI after SICF in terms of depth of semen deposition, time to pass the cervix, PR and LR (p < 0.05). In conclusion, the use of a catheter that allowed transcervical uterine deposition of semen without excessive manipulation led to satisfactory pregnancy rates.

Determination of minimum alveolar concentration and cardiovascular effects of desflurane in positive-pressure ventilated sheep.

OBJECTIVE To determine the minimum alveolar concentration of desflurane (MACDES) and effects on cardiovascular variables in positive-pressure ventilated sheep. ANIMALS 13 adult female sheep. PROCEDURES Anesthesia was induced with desflurane. After a 30-minute equilibration at an end-tidal concentration of desflurane (etDES) of 10.5%, an electrical stimulus (5 Hz/ms and 50 mA) was applied for 1 minute or until gross purposeful movement occurred. The etDES was then changed by 0.5% (modified up-down method), depending on whether a positive motor response had been elicited, and stimulation was repeated. The MACDES was the etDES midway between a positive and negative response. After MACDES was determined, etDES was increased to 1.3 and 1.6 MACDES. Animals were allowed to equilibrate for 15 minutes, and cardiovascular, blood gas, acid-base, and hematologic variables were measured. Times to induction of anesthesia, extubation, attainment of sternal position, and standing and duration of anesthesia were recorded. RESULTS Mean ± SD MACDES was 9.81 ± 0.79%. Times to intubation, extubation, and standing were 4.81 ± 2.21 minutes, 14.09 ± 4.05 minutes, and 32.4 ± 12.5 minutes, respectively. Duration of anesthesia was 226 ± 22 minutes. Heart rate increased significantly at induction of anesthesia but otherwise remained at preanesthetic rates. Arterial blood pressures progressively decreased with increasing etDES; pressures increased slightly only in response to noxious stimulation. CONCLUSIONS AND CLINICAL RELEVANCE The MACDES determined here compared favorably with that determined for other sheep populations and indicated similar anesthetic potency as in other species. Desflurane caused dose-dependent arterial hypotension, which indicated the need for careful blood pressure monitoring.

Determination of the minimum alveolar concentration (MAC) and cardiopulmonary effects of sevoflurane in sheep.

OBJECTIVE: To determine sevoflurane's minimum alveolar concentration (MACSEVO) and its cardiopulmonary effects in sheep. STUDY DESIGN: Prospective experimental study. ANIMALS: A group of 10 female nonpregnant Sardinian milk sheep. METHODS: Anesthesia was induced in each sheep twice with sevoflurane in oxygen. After a 30 minute equilibration at end-tidal sevoflurane concentration (Fe'Sevo) of 2.8%, an electrical stimulus (5 Hz/1 ms/50 mA) was applied to the right thoracic limb for 1 minute or until gross purposeful movement occurred. The Fe'Sevo was then changed using a 0.2% up-and-down protocol, dependent on whether or not the response was positive, and then noxious stimulation was repeated. The MACSEVO was defined as the mean Fe'Sevo between that allowing purposeful movement and that not. The group of 10 sheep were re-anesthetized and MACSEVO was re-determined. Thereafter, Fe'Sevo was maintained for 15 minutes each at concentrations corresponding to 1.0, 1.3, 1.6, 1.9 and 0.75 MACSEVO multiples, and cardiopulmonary, blood gas, acid-base variables and plasma electrolytes were determined. Also, time to induction of anesthesia, extubation and recovery were recorded. RESULTS: The mean ± standard deviation of the MACSEVO was 2.74 ± 0.38%. Median (interquartile range) time to intubation was 3.13 (2.98-3.33) minutes, time to extubation was 6.85 ± 2.65 minutes and time to recovery was 13.4 ± 5.2 minutes. With increasing Fe'Sevo, arterial blood pressures progressively decreased as did minute ventilation, which in turn caused end-tidal carbon dioxide, arterial partial pressure of carbon dioxide and bicarbonate values to steadily increase without significantly affecting arterial partial pressure of oxygen. CONCLUSIONS AND CLINICAL RELEVANCE: The reported MACSEVO agrees with published data in this and other species. Administration of sevoflurane in sheep caused marked hemodynamic and respiratory depression, but soon after turning off the vaporizer, sheep could be extubated and recovered rapidly and event-free.

MAC-sparing effect of nitrous oxide in sevoflurane anesthetized sheep and its reversal with systemic atipamezole administration.

INTRODUCTION: Nitrous oxide (N2O) is an anesthetic gas with antinociceptive properties and reduces the minimum alveolar concentration (MAC) for volatile anesthetic agents, potentially through mechanisms involving central alpha2-adrenoceptors. We hypothesized that 70% N2O in the inspired gas will significantly reduce the MAC of sevoflurane (MACSEVO) in sheep, and that this effect can be reversed by systemic atipamezole. MATERIALS AND METHODS: Animals were initially anesthetized with SEVO in oxygen (O2) and exposed to an electrical current as supramaximal noxious stimulus in order to determine MACSEVO (in duplicates). Thereafter, 70% N2O was added to the inspired gas and the MAC re-determined in the presence of N2O (MACSN). A subgroup of sheep were anesthetized a second time with SEVO/N2O for re-determination of MACSN, after which atipamezole (0.2 mg kg-1, IV) was administered for MACSNA determinations. Sheep were anesthetized a third time, initially with only SEVO/O2 to re-determine MACSEVO, after which atipamezole (0.2 mg kg-1, IV) was administered for determination of MACSA. RESULTS: MACSEVO was 2.7 (0.3)% [mean (standard deviation)]. Addition of N2O resulted in a 37% reduction of MACSEVO to MACSN of 1.7 (0.2)% (p <0.0001). Atipamezole reversed this effect, producing a MACSNA of 3.1 (0.7)%, which did not differ from MACSEVO (p = 0.12). MACSEVO did not differ from MACSA (p = 0.69). Cardiorespiratory variables were not different among experimental groups except a lower ETCO2 in animals exposed to SEVO/N2O. CONCLUSIONS: N2O produces significant MACSEVO-reduction in sheep; this effect is completely reversed by IV atipamezole confirming the involvement of alpha2-adrenoreceptors in the MAC-sparing action of N2O.

Relationships among neuroscore, magnetic resonance imaging features, and intracranial pressure in sheep affected by slow-growing brain lesions.

Diagnosing high intracranial pressure by clinical and diagnostic imaging is particularly challenging for chronic or slow-growing lesions. The aim of this prospective case-control study is to determine whether the neuroscore and brain magnetic resonance imaging (MRI) are related to the direct measurement of intracranial pressure in sheep affected by intracranial slow-growing lesions due to chronic cerebral coenurosis (Coenurus cerebralis). Seventeen affected and 10 control sheep were included. All animals underwent a neurological examination, MRI of the brain, and direct measurement of intracranial pressure. The severity of clinical signs and MRI findings were scored. Data were statistically analyzed. The invasive intracranial pressure value was higher in affected animals. A severely altered neuroscore is related to an increased intracranial pressure beyond the normal threshold (P < 0.05). The volume of the calvarium was larger in affected animals than in control animals (P = 0.0001) and was positively influenced by the presence and volume of the parasitic cyst (r = 0.7881, P < 0.01). Several degrees of deviation and deformation of both the ventricular system and brain parenchyma were detected by MRI. Subjective MRI findings were not associated with intracranial hypertension. In conclusion, this study shows that in sheep affected by slow-growing lesions, severe alterations in the neuroscore and the results of objective MRI are related to an increased intracranial pressure beyond the normal threshold.

Clinical evolution of cerebral coenurosis from invasive to chronic infection in sheep and a goat.

This survey describes the evolution in vivo of Coenurus cerebralis in small ruminants. At presentation, neurological signs and cerebrospinal fluid (CSF) features were suggestive of multifocal or diffuse inflammatory reaction. Magnetic resonance imaging (MRI) captured the transition between the invasive and quiescent phase of the infection, revealing the concurrent presence of meningitis and small cysts. During the quiescent phase, in all animals, neurological symptoms disappeared, and cerebrospinal fluid was unremarkable while cysts grew progressively. Subsequently, the onset of neurological symptoms coincided with MRI signs of diffuse or localized increase of intracranial pressure, as confirmed by direct intracranial pressure measuring. All the animals had an excellent post-surgical recovery. This is the first report describing the evolution of coenurosis in vivo. Sequential imaging allowed describing interesting such as the death of some coenuri and different parasite growth rate in the same host.