Utilization of substrates for testosterone and estradiol-17β production by small follicles of the chicken ovary

A study was conducted to characterize steroidogenesis in small ovarian follicles (1–10 mm in diameter) of the hen. The aims of our study were: 1) to determine basal estradiol-17β (E₂) production by different sizes of small follicles; 2) to determine the ability of intact small follicles to utilize exogenous substrates for testosterone (T) and E₂ production; and 3) to investigate the preferred steroidogenic pathway in small follicles. Small follicles which had not entered the hierarchy were isolated from ovaries obtained 2 hr after oviposition and divided into three groups: small white follicles (SWF; 1–2 mm in diameter), large white follicles (LWF; 2–4 mm in diameter), and small yellow follicles (SYF; 5–10 mm in diameter). Yolk and granulosa cells were removed from LWFs and SYFs and the remaining theca layer was called a follicle shell. Intact follicles or follicle shells (4/4 ml/tube) were incubated in avian Ringer's buffer supplemented with 10 mM HEPES and 0.1% BSA at 37°C for 3 hr with various treatments. Testosterone and E₂ were measured in the medium. The SYFs and their corresponding follicle shells produced the greatest amount of E₂ when E₂ production was expressed per follicle. Addition of 2 mM 8-Br-cAMP to the incubation medium stimulated E₂ production by all sizes of follicles and follicle shells. However, follicle shells produced lower basal- and 8-Br-cAMP-stimulated E₂ secretion compared to corresponding intact follicles. There was no significant difference in E₂ production in response to various concentrations of 25-hydroxycholesterol (25-OH-CHOL; 0–100 μM) by intact follicles and follicle shells. On the contrary, intact follicles and follicle shells produced T and E₂ in a dose-dependent manner in response to increasing concentrations (0–100 μM) of pregnenolone (P₅). Intact follicles also used progesterone (P₄) and dehydroepiandrosterone (DHEA) as substrates for T and E₂ production. DHEA was the preferred substrate for steroid production compared to P₄. In summary, we found that: 1) steroidogenesis in small follicles is regulated by a cAMP-dependent protein kinase A second messenger system; 2) adequate amounts of endogenous cholesterol are available for steroidogenesis; and 3) both Δ⁵ and Δ⁴ pathways are functional in small follicles and the Δ⁵ pathway may be the preferred steroidogenic pathway. pathway.     

Effect of body position on the arterial partial pressures of oxygen and carbon dioxide in spontaneously breathing,conscious dogs in an intensive care unit

Objective – To evaluate the effect of body position on the arterial partial pressures of oxygen and carbon dioxide (PaO₂, PaCO₂), and the efficiency of pulmonary oxygen uptake as estimated by alveolar‐arterial oxygen difference (A‐a difference). Design – Prospective, randomized, crossover study. Setting – University teaching hospital, intensive care unit. Animals – Twenty‐one spontaneously breathing, conscious, canine patients with arterial catheters placed as part of their management strategy. Interventions – Patients were placed randomly into lateral or sternal recumbency. PaO₂ and PaCO₂ were measured after 15 minutes in this position. Patients were then repositioned into the opposite position and after 15 minutes the parameters were remeasured. Measurements and Main Results – Results presented as median (interquartile range). PaO₂ was significantly higher (P=0.001) when patients were positioned in sternal, 91.2 mm Hg (86.0–96.1 mm Hg), compared with lateral recumbency, 86.4 mm Hg (73.9–90.9 mm Hg). The median change was 5.4 mm Hg (1.1–17.9 mm Hg). All 7 dogs with a PaO₂<80 mm Hg in lateral recumbency had improved arterial oxygenation in sternal recumbency, median increase 17.4 mm Hg with a range of 3.8–29.7 mm Hg. PaCO₂ levels when patients were in sternal recumbency, 30.5 mm Hg (27.3–32.7 mm Hg) were not significantly different from those in lateral recumbency, 32.2 mm Hg (28.3–36.0 mm Hg) (P=0.07). The median change was ?1.9 mm Hg (?3.6–0.77 mm Hg). A‐a differences were significantly lower (P=0.005) when patients were positioned in sternal recumbency, 21.7 mm Hg (17.3–27.7 mm Hg), compared with lateral recumbency, 24.6 mm Hg (20.4–36.3 mm Hg). The median change was ?3.1 mm Hg (?14.6–0.9 mm Hg). Conclusions – PaO₂ was significantly higher when animals were positioned in sternal recumbency compared with lateral recumbency, predominantly due to improved pulmonary oxygen uptake (decreased A‐a difference) rather than increased alveolar ventilation (decreased PaCO₂). Patients with hypoxemia (defined as PaO₂<80 mm Hg) in lateral recumbency may benefit from being placed in sternal recumbency. Sternal recumbency is recommended to improve oxygenation in hypoxemic patients.     

Impact of endotracheal tube size and cuff pressure on tracheal and laryngeal mucosa of adult horses

ObjectiveTo assess the effects of two sizes of silicone endotracheal tubes with internal diameter 26 mm (ETT₂₆) and 30 mm (ETT₃₀) inflated to minimum occlusive volume on tracheal and laryngeal mucosa of adult horses anesthetized for 2 hours with isoflurane.Study designProspective, randomized, blinded, crossover experimental study.AnimalsA total of eight healthy adult mares.MethodsUpper airway endoscopy and ultrasound measurements of internal tracheal diameter were performed the day before anesthesia. Horses were anesthetized and orotracheally intubated with ETT₂₆ or ETT₃₀. Ease of intubation was scored. The cuff was inflated in 10 mL increments to produce a seal. Final volume of air used and intracuff (IC) pressure (measured by pressure transducer) were recorded. At the end of anesthesia, a manometer was used to measure IC pressure and these measurements compared against measurements from the pressure transducer. Laryngeal and tracheal mucosa were assessed via endoscopy and assigned a score 0–3 before anesthesia, and at 2 and 24 hours following extubation.ResultsData are from seven horses because one horse with laryngeal hemiplegia was excluded. Mean tracheal ultrasound measurement was 3.5 ± 0.4 cm. No significant differences were noted between endotracheal tube sizes for intubation score, IC pressures, inflation volumes or tracheal or laryngeal injury scores at any time point. IC pressure measured by manometer was slightly higher than that by transducer (+1.0 ± 2.8 mmHg).Conclusions and clinical relevanceResults identified no clear advantage of one endotracheal tube size over the other in the population of horses studied, when endotracheal intubation is properly applied and IC pressure is carefully monitored. However, given that ETT₂₆ was associated with the highest observed IC pressures and the only observed incidents of tracheal circumferential erythema, the larger ETT₃₀ may be the better choice in most cases where tracheal size is sufficient.     

Comparison of mainstream (Capnostat 5) and two low-flow sidestream capnometers (VM-2500-S and Capnostream) in spontaneously breathing rabbits anesthetized with a Bain coaxial breathing system

ObjectiveTo evaluate agreement with PaCO₂ of two low sampling rate sidestream capnometers and a mainstream capnometer in rabbits and the effect of using high fresh gas flow from a Bain coaxial breathing system.Study designProspective, crossover study.AnimalsA total of 10 New Zealand White rabbits weighing 3.4 ± 0.3 kg [mean ± standard deviation (SD)].MethodsTwo sidestream analyzers (Viamed VM-2500-S and Capnostream 35) with a sampling rate of 50 mL minute^–1 and a mainstream capnometer (Capnostat 5) were tested. All capnometers used infrared spectroscopy and advanced microprocessor technology. Rabbits were anesthetized and intubated with noncuffed endotracheal tubes of 3 mm internal diameter and adequate seal. A sidestream sampling adapter or the mainstream capnometer was attached to the endotracheal tube and connected to a Bain coaxial breathing system. Oxygen (1.5 L minute^–1) delivered sevoflurane to maintain anesthesia. An auricular artery catheter allowed blood sampling for PaCO₂ analysis corrected to rectal temperature. Inspired and end-tidal carbon dioxide (Pe′CO₂) measurements were recorded during blood sample withdrawal. From each rabbit, 10 paired PaCO₂/Pe′CO₂ measurements were obtained. Each rabbit was recovered from anesthesia and was anesthetized again with an alternate capnometer after 1 week. Data were analyzed using Bland–Altman and two-way anova for repeated measures.ResultsAnalysis included 100 paired samples. Negative bias reflects underestimation of PaCO₂. Bland–Altman mean (±1.95 SD) was –16.7 (–35.2 to 1.8) mmHg for Capnostat 5, –27.9 (–48.6 to –7.2) mmHg for Viamed, and –18.1 (–34.3 to –1.9) mmHg for Capnostream. Viamed PaCO₂–Pe′CO₂ gradient was greater than other two capnometers.ConclusionsAll three capnometers underestimated PaCO₂. Capnostat 5 and Capnostream performed similarly.Clinical relevanceThese capnometers underestimated PaCO₂ in spontaneously breathing rabbits anesthetized using a Bain coaxial breathing system with high fresh gas flows.     

Comparison between mainstream (Capnostat 5) and a low-flow sidestream capnometer (Capnostream) in mechanically ventilated,sevoflurane-anesthetized rabbits using a Bain coaxial delivery system

ObjectiveTo evaluate agreement between end-tidal carbon dioxide (Pe′CO₂) and PaCO₂ with sidestream and mainstream capnometers in mechanically ventilated anesthetized rabbits, with two ventilatory strategies.Study designProspective experimental study.AnimalsA total of 10 New Zealand White rabbits weighing 3.6 ± 0.3 kg (mean ± standard deviation).MethodsRabbits anesthetized with sevoflurane were intubated with an uncuffed endotracheal tube (3.0 mm internal diameter) and adequate seal. For Pe′CO₂, the sidestream capnometer sampling adapter or the mainstream capnometer was placed between the endotracheal tube and Bain breathing system (1.5 L minute^–1 oxygen). PaCO₂ was obtained from arterial blood collected every 5 minutes. A time-cycled ventilator delivered an inspiratory time of 1 second and 12 or 20 breaths minute^–1. Peak inspiratory pressure was initially set to achieve Pe′CO₂ normocapnia of 35–45 mmHg (4.6–6.0 kPa). A total of five paired Pe′CO₂ and PaCO₂ measurements were obtained with each ventilation mode for each capnometer. Anesthetic episodes were separated by 7 days. Agreement was assessed using Bland-Altman analysis and linear mixed models; p < 0.05.ResultsThere were 90 and 83 pairs for the mainstream and sidestream capnometers, respectively. The mainstream capnometer underestimated PaCO₂ by 12.6 ± 2.9 mmHg (proportional bias 0.44 ± 0.06 mmHg per 1 mmHg PaCO₂ increase). With the sidestream capnometer, ventilation mode had a significant effect on Pe′CO₂. At 12 breaths minute^–1, Pe′CO₂ underestimated PaCO₂ by 23.9 ± 8.2 mmHg (proportional bias: 0.81 ± 0.18 mmHg per 1 mmHg PaCO₂ increase). At 20 breaths minute^–1, Pe′CO₂ underestimated PaCO₂ by 38.8 ± 5.0 mmHg (proportional bias 1.13 ± 0.10 mmHg per 1 mmHg PaCO₂ increase).Conclusions and clinical relevanceBoth capnometers underestimated PaCO₂. The sidestream capnometer underestimated PaCO₂ more than the mainstream capnometer, and was affected by ventilation mode.     

Potential risks,prognostic indicators,and diagnostic and treatment modalities affecting survival in dogs with presumptive aspiration pneumonia: 125 cases (2005–2008)

Objective – To evaluate a clinical population of dogs diagnosed with presumptive aspiration pneumonia (AP) and determine diagnostic and treatment modalities contributing to survival. Design – Retrospective study. Setting – A university veterinary teaching hospital in an urban setting. Animals – One hundred and twenty‐five dogs with presumed AP treated from 2005 to 2008. Interventions – None. Measurements and Main Results – Dogs with presumptive AP identified by a review of medical records had an overall survival of 81.6% (102/125). Male large‐breed dogs (mean 24.9 kg; 82/125) were overrepresented and were more likely to develop AP in this study population. Recent anesthesia had been performed in 16% (20/125), and vomiting was reported in 64% (80/125). The most common radiographic findings were a predominantly alveolar pattern (187/272, [68.8%] total lung lobes) in the right middle lung lobe (80/115, [69.6%]). A mean of 2 lung lobes were involved radiographically, and the relationship between survival and the number of lung lobes affected was statistically significant (P=0.04). Neutrophilia with a left shift was common with no significant change on consecutive daily evaluations. The mean PaO₂ was 77.7 mm Hg (SD, 17.5 mm Hg) (range, 40.7–100 mm Hg) with a median alveolar‐arterial gradient of 41.1 mm Hg (range, 8.1–81.8 mm Hg). In this study population, 37.6% (47/125) of dogs had microbial cultures performed and of these, 76.6% (36/47) were positive for growth; Escherichia coli (38.8%), Mycoplasma spp. (21.3%), Pasturella spp. (19.1%), and Staphylococcus spp. (17%) were the most common isolates in either single or multiagent infections. No treatment modality was statistically associated with increased survival. Colloid therapy was a negative prognostic indicator. Conclusions – In this study the overall prognosis for AP was good. Patients with only 1 affected lung lobe appeared more likely to survive. Supportive treatment modalities are warranted for the hospitalized patient, although no individual treatment method was found to be clearly superior to others.     

Endotracheal intubation in horses: a study of two cuff inflation pressures, correlation with liquid aspiration, and tracheal wall damage

Nine adult horses were anesthetized for a nonsurvival abdominal adhesion study. Horses were randomly assigned into two groups to receive endotracheal tube cuff pressures of either 80 cm H₂O (Group P80) or 120 cm H₂O (Group P120). After intubation (Bivona 30 mm ID), anesthesia was maintained with isoflurane. Horses were ventilated 10 times per minute with a suitable inspiratory pressure to maintain Pe ′CO₂ in the 35–40 mm Hg (4.7–6.0 kPa) range. Cuff pressure was continuously monitored with a pressure transducer (TruWave, Baxter) calibrated to the atmospheric pressure and maintained at a constant pressure. Twenty‐five millilitres of methylene blue dye in saline were instilled proximal to the cuff over 5 minutes. The horses were euthanized 123 ± 23 minutes later (mean ± SD). Immediately, the trachea was opened distal to the tip of the endotracheal tube, and the mucosa was observed for evidence of dye leaking past the cuff. The cervical trachea was resected and the lumen exposed by a ventral longitudinal incision. Biopsies (1–2 rings) were obtained at mid‐cuff level and distal to the tip of the endotracheal tube, and placed in formalin for later histologic examinations (H&E stain). Methylene blue stain was not observed distal to the endotracheal tube cuff in any horse. Visual examination of the tracheal mucosa revealed hyperemic or hemorrhagic lesions at the level of cuff contact both ventrally and dorsally. Histologic changes included epithelium damage, submucosal neutrophil infiltrates, and acute submucosal hemorrhages. P80 horses had none or focal to multifocal lesions on the ventral and dorsal aspects of the rings. P120 horses had multifocal to diffuse lesions on all aspects (dorsal, ventral, and lateral). We concluded that the endotracheal tube cuff produced a seal sufficient to prevent leakage at both pressures. Tracheal damages on gross and microscopic examinations were more severe and occurred more frequently at the higher cuff pressure.     

Aerosolized salbutamol (albuterol) improves PaO2 in hypoxaemic anaesthetized horses – a prospective clinical trial in 81 horses

Objective To compare the arterial pH and blood gas values, heart rate and mean arterial blood pressure, in hypoxaemic anaesthetized horses, before and after treatment, with a salbutamol (albuterol) aerosol. Animal population Eighty‐one client‐owned horses weighing between 114 and 925 kg. Fifty‐seven underwent emergency abdominal surgery and 24 were anaesthetized for elective procedures. Materials and methods Pre‐anaesthetic medication included xylazine, detomidine, butorphanol and morphine, alone or in various combinations. Induction of anaesthesia was achieved with guaifenesin and ketamine, diazepam and ketamine, or guaifenesin and thiopental. The trachea of all animals was intubated and anaesthesia maintained with either halothane (33 horses) or isoflurane (48 horses) in oxygen. Heart rate and rhythm were monitored continuously. Arterial blood pressure was monitored directly, and arterial blood collected for pH and blood gas analyses. When arterial PaO₂ fell below 9.3 kPa (70 mm Hg) and failed to respond to corrective measures including positive pressure ventilation and treatment of hypotension (mean arterial blood pressures <70 mm Hg), a salbutamol aerosol (2 µg kg^?1) was delivered via the endotracheal tube. Twenty minutes later, a second arterial blood sample was analysed. Results There were no significant differences in mean arterial blood pressure, heart rate, arterial pH, base excess and bicarbonate before and after treatment. Arterial O₂ tension increased significantly from a mean ± SD of 8.3 ± 1.7 kPa (62.4 ± 13.1 mm Hg) before administration to 15.9 ± 9.8 kPa (119.4 ± 57.7 mm Hg) after treatment. There was a small but significant decrease in PaCO₂ from 7.4 ± 1.5 kPa (55.2 ± 11.2 mm Hg) to 7.0 ± 1.3 kPa (52.9 ± 9.8 mm Hg) between sample times. No changes in heart rhythm were observed. A high percentage (approximately 70%) of animals sweated following treatment. Conclusions Salbutamol administered at a dose of 2 µg kg^?1 via the endotracheal tube of anaesthetized horses with PaO₂ values less than 9.3 kPa (70 mm Hg) resulted in an almost two‐fold increase in PaO₂ values within 20 minutes of treatment. No changes in heart rate or mean arterial blood pressure were associated with the use of salbutamol in this study. The improvement in PaO₂ may be a result of bronchodilatation and improved ventilation, increased perfusion secondary to an increase in cardiac output, or a combination of these two factors. Cardiac output and ventilation–perfusion distribution were not measured in this study; therefore, the reason for the increase in PaO₂ values cannot be conclusively determined. Clinical relevance Administration of a salbutamol aerosol is a simple but effective technique that can be used to improve PaO₂ values in hypoxaemic horses during inhalant anaesthesia with no apparent detrimental side effects.     

T2‐WEIGHTED MAGNETIC RESONANCE IMAGING MEASUREMENTS OF OPTIC NERVE SHEATH DIAMETER IN DOGS WITH AND WITHOUT PRESUMED INTRACRANIAL HYPERTENSION

Intracranial hypertension is a cause of cerebral ischemia and neurologic deficits in dogs. Goals of this retrospective study were to test interobserver agreement for MRI measurements of optic nerve sheath diameter and associations between optic nerve sheath diameter, signalment data, and presumed intracranial hypertension status in a cohort of dogs. A veterinary radiologist interpreted scans of 100 dogs and dogs were assigned to groups based on presence or absence of at least two MRI characteristics of presumed intracranial hypertension. Two observers who were unaware of group status independently measured optic nerve diameter from transverse T2‐weighted sequences. Mean optic nerve sheath diameter for all dogs was 3 mm (1–4 mm). The mean difference between observers was 0.3 mm (limits of agreement, ?0.4 and 1.0 mm). There was no correlation between optic nerve sheath diameter and age for either observer (r = ?0.06 to 0.00) but a moderate positive correlation was observed between optic nerve sheath diameter and body weight for both observers (r = 0.70–0.76). The 22 dogs with presumed intracranial hypertension weighed less than the 78 dogs without (P = 0.02) and were more often female (P = 0.04). Dogs with presumed intracranial hypertension had a larger ratio of optic nerve sheath diameter to body weight for each observer‐side pair (P = 0.01–0.04) than dogs without. Findings indicated that the ratio of MRI optic nerve sheath diameter relative to body weight may be a repeatable predictor of intracranial hypertension in dogs.     

Measurement of tracheal diameters using ultrasonography versus computed tomography in Beagle dogs

ObjectiveTo compare ultrasonography with computed tomography (CT) for assessment of tracheal diameter as a feasibility study for endotracheal tube selection.Study designProspective study.AnimalsA total of nine Beagle dogs with a median (interquartile range) weight of 7.4 (7.2–7.7) kg.MethodsTracheal diameter measurements were obtained at two locations: 1 cm proximal to caudal border of the cricoid cartilage (sublaryngeal; SL) and dorsal to above cranial border of the manubrium (thoracic inlet; TI). For CT, dogs were anesthetized with propofol and sevoflurane, in sternal recumbency, and measurements obtained after controlled ventilation–induced apnea and the endotracheal tube cuff was deflated. Transverse diameter, right and left 45° oblique diameters were measured. For ultrasonography, unsedated dogs were standing with slight neck extension, and images obtained in ventrodorsal, 45° right and left oblique ways after expiration. Diameters between the tracheal lumen mucosal borders were measured. The degree of agreement between the tracheal diameters measured at SL and TI locations with CT (TD_CT-SL and TD_CT-TI) and ultrasonography (TD_US-SL and TD_US-TI) was verified using the Bland-Altman method.ResultsThe agreement between the measurements obtained with CT and ultrasonography was revealed by Bland-Altman analyses, although ultrasonography tended to slightly underestimate the tracheal diameter.Conclusions and clinical relevanceUltrasonography can be applied for tracheal diameter measurement. Although further studies are required, an endotracheal tube selection method, using ultrasonography, could be proposed.     

Pharmacokinetics of tildipirosin in porcine plasma,lung tissue,and bronchial fluid and effects of test conditions on in vitro activity against reference strains and field isolates of Actinobacillus pleuropneumoniae

Rose, M, Menge, M, Bohland, C, Zschiesche, E, Wilhelm, C, Kilp, S, Metz, W, Allan, M, Röpke, R, Nürnberger, M Pharmacokinetics of tildipirosin in porcine plasma, lung tissue, and bronchial fluid and effects of test conditions on in vitro activity against reference strains and field isolates of Actinobacillus pleuropneumoniae. J. vet. Pharmacol. Therap.  36 , 140–153. The pharmacokinetics of tildipirosin (Zuprevo^® 40 mg/mL solution for injection for pigs), a novel 16‐membered‐ring macrolide for the treatment for swine respiratory disease (SRD), was investigated in studies collecting blood plasma and postmortem samples of lung tissue and bronchial fluid (BF) from swine. In view of factors influencing the in vitro activity of macrolides, and for the interpretation of tildipirosin pharmacokinetics in relation to minimum inhibitory concentrations (MIC), additional experiments were conducted to study the effects of pH, carbon dioxide‐enriched atmosphere, buffers, and serum on tildipirosin MICs for various reference strains and Actinobacillus (A.) pleuropneumoniae field isolates. After single intramuscular (i.m.) injection at 4 mg/kg body weight, maximum plasma concentration (C_max) was 0.9 μg/mL observed within 23 min (T_max). Mean residence time from the time of dosing to the time of last measurable concentration (MRT_last) and terminal half‐life (T_1/2) both were about 4 days. A dose–response relationship with no significant sex effect is observed for area under the plasma concentration–time curve from time 0 to the last sampling time with a quantifiable drug concentration (AUC_last) over the range of doses up to 6 mg/kg. However, linear dose proportionality could not be proven with statistical methods. The time–concentration profile of tildipirosin in BF and lung far exceeded that in blood plasma. In lung, tildipirosin concentrations reached 3.1 μg/g at 2 h, peaked at 4.3 μg/g at day 1, and slowly declined to 0.8 μg/g at day 17. In BF, tildipirosin levels were 14.3, 7.0, and 6.5 μg/g at days 5, 10, and 14. T_1/2 in lung was ～7 days. Tildipirosin is rapidly and extensively distributed to the respiratory tract followed by slow elimination. Culture media pH and carbon dioxide‐enriched atmosphere (CO₂‐EA) had a marked impact on in vitro activity of tildipirosin in reference strains of various rapidly growing aerobic and fastidious bacteria including Histophilus (H.) somni ATCC 700025 and A. pleuropneumoniae ATCC 27090. For A. pleuropneumoniae ATCC 27090 testing conditions without CO₂‐EA resulted in reduced acidification of culture media pH and a reduction in the minimum inhibitory concentrations compared to standard in vitro test conditions by 2 log₂ dilution steps (4‐fold) from 8 to 2 μg/mL. Supplementary buffering of standard culture media resulted in a reduction in the A. pleuropneumoniae (n = 8) MIC range by 4 log₂ dilution steps (16‐fold) from 8–16 to 0.5–1 μg/mL. Incremental supplementation of culture media with 50% serum resulted in noticeable shifts to lower minimum or maximum MICs by at least 2 log₂ dilution steps (≥4‐fold) in all aerobic and fastidious reference strains tested except for Pasteurella (P.) multocida. The MIC of A. pleuropneumoniae ATCC 27090 decreased by 2–4 log₂ dilution steps (4 to 16‐fold) from 8 to 0.5–2 μg/mL when 50% serum was added to the standard assay. Considering a higher presence of serum and the rather neutral pH conditions maintained in vivo, it is suggested to take the influence of these factors on in vitro activity into account when interpreting tildipirosin MICs for A. pleuropneumoniae in relation to pharmacokinetics.     

THE VETMOUSETRAP™: A DEVICE FOR COMPUTED TOMOGRAPHIC IMAGING OF THE THORAX OF AWAKE CATS

The VetMousetrap^?, a novel device that allows computed tomography (CT) of awake cats and provides a clinically supportive environment, is described. Ten normal cats were used to test the device for ambient internal oxygen, carbon dioxide levels, and temperature. Twenty‐two awake normal cats were imaged using a 16‐multi‐slice helical CT unit to evaluate dose‐equivalent protocols. Two different X‐ray tube potentials (kV), 80 and 120, and two different helical pitches, 0.562 and 1.75, were evaluated. The signal intensity of the pulmonary parenchyma (SI_lung), signal intensity of background (SI_backgr), contrast, noise, signal‐to‐noise ratio (SNR), and contrast‐to‐noise ratio (CNR) were calculated. Three evaluators ranked the images for sharpness of liver margins, motion, helical, and windmill artifacts. CT was successfully completed in 20 of 22 cats. No artifacts directly related to the device were detected. Overall, 75 of 80 (94%) examinations were judged to have absent or minimal motion artifact. A statistically significant difference was found for SNR (P=0.001) and CNR (P=0.001) between all protocols. The higher pitch protocols had significantly lower noise and higher SNR and CNR, lower motion artifact but greater helical artifacts. A protocol using 80 kV, 130 mA, 0.5 s, and 0.562 pitch with 1.25 mm slice thickness, and 0.625 mm slice reconstruction interval is recommended. The VetMousetrap^? appears to provide the opportunity for diagnostic CT imaging of the thorax of awake cats.     

Clinical evaluation of balanced anesthesia using infusion of midazolam–ketamine–medetomidine in combination with inhalation of sevoflurane (MKM–OS anesthesia) in horses

MKM–OS anesthesia provides general anesthesia with minimum cardiovascular depression in experimental horses. The purpose of this study was to evaluate the effect of MKM–OS anesthesia in clinical cases. Sixty‐eight horses were anesthetized with MKM–OS anesthesia for selective or emergency surgery. The horse physical status was categorized based upon the American Society of Anesthesiologists (ASA) classification scheme. Forty‐four horses were classified as ASA I or II (low‐risk; 30 soft tissue, eight ophthalmic, and six orthopedic surgeries) and 24 horses were classified as ASA III to V (high‐risk; 24 emergency colic surgeries). All horses were administered medetomidine (0.005 mg kg^–1 IV) as premedication and anesthetized with ketamine (2.5 mg kg^–1 IV) and midazolam (0.04 mg kg^–1 IV). The horses were orotracheally intubated and connected to a large animal breathing circuit that delivered oxygen‐sevoflurane and administered the midazolam (0.8 mg mL^–1)‐ketamine (40 mg mL^–1)‐medetomidine (0.05 mg mL^–1) drug combination at a rate of 0.025 mL kg^–1 hour^–1. Surgical anesthesia was maintained by controlling the dial setting of the sevoflurane vaporizer and achieved by delivering 1.6–1.8% of end‐tidal sevoflurane concentration. All horses were mechanically ventilated during anesthesia. Hypercapnia and hypoxia were not sufficiently improved in high‐risk horses (PaCO₂; low‐risk 45–53 mm Hg versus high‐risk 56–60 mm Hg, p < 0.01: PaO₂ low‐risk 248–388 mm Hg versus high‐risk 95–180 mm Hg, p < 0.01). Heart rate was significantly higher in high‐risk horses (low‐risk 37–42 bpm versus high‐risk 44–73 bpm, p < 0.01). Dobutamine infusion was required in five low‐risk (11%) and 17 high‐risk horses (68%) to maintain mean arterial blood pressure >70 mm Hg. Eleven high‐risk horses died during the perioperative period (three euthanized during surgery, two died during recovery, six died after recovery). The quality of recovery was good in low‐risk horses and good to satisfactory in high‐risk horses. MKM–OS anesthesia provided excellent surgical anesthesia with minimal to mild cardiovascular depression in low risk‐horses and mild to moderate cardiovascular depression in high risk‐horses. The possibility of preserve cardiovascular function could be the advantage of MKM–OS anesthesia in high‐risk horses.     

Protein-losing enteropathy in dogs is associated with decreased fecal proteolytic activity

Background: Measurement of proteolytic activity in feces is a traditional method for the diagnosis of exocrine pancreatic insufficiency (EPI). A drawback of this method is the occurrence of falsely low results that may lead to a false‐positive diagnosis of EPI. We hypothesized that intestinal loss of serum proteinase inhibitors in protein‐losing enteropathy (PLE) may inhibit fecal proteolytic activity and be a potential source of false low results. Objective: The objective of this study was to determine the effect of PLE on fecal proteolytic activity in dogs. Methods: Fecal proteolytic activity was measured using a radial diffusion casein digestion assay in 12 samples from 4 clinically healthy control dogs and 30 samples from 16 dogs with PLE. Gastrointestinal protein loss was assessed using an ELISA to determine fecal canine α₁‐proteinase inhibitor concentration. The relationship between the concentration of canine α₁‐proteinase inhibitor in the feces and the diameter cleared in the casein digestion assay was determined. The mean clearing diameter was compared between control dogs and dogs with PLE. Results: A significant negative correlation was observed between fecal canine α₁‐proteinase inhibitor concentration and casein clearing diameter (P < .001, Pearson r=—.6317, r 2 =.3999). Mean clearing diameter was significantly lower in dogs with PLE than in control dogs (12.63 vs 16.83 mm, P < .001, two‐tailed Student's t‐test). Conclusion: Increased fecal loss of α₁‐proteinase inhibitor in dogs with PLE is associated with a significant decrease in fecal proteolytic activity and may result in a false positive diagnosis of EPI.     

Effect of Follicle Size on In Vitro Maturation of Pre‐Pubertal Porcine Cumulus Oocyte Complexes

Very small follicles (<3.0 mm diameter) are over‐represented on the surface of ovaries of non‐cycling pigs, and the oocytes collected from these follicles generally have reduced developmental competence in vitro. This study examined the effect of follicle size on the nuclear maturation (n = 608), the potential of parthenogenetic activation (n = 243) and the cyclic AMP (cAMP) content of pre‐pubertal porcine oocytes (n = 480). In addition, the influence of follicle size on steroid hormone synthesis was analysed. Cumulus oocyte complexes (COCs) flushed from small (2.5–4.0 mm) or large (4.5–6.0 mm) ovarian follicles were cultured for 0, 28 and 46 h. After 46 h of IVM, a greater proportion of oocytes from 4.5‐ to 6.0‐mm follicles reach metaphase II (MII) compared with those from follicles with 2.5–4.0 mm of diameter (96.1 vs 77.0%, respectively; p < 0.001). Parthenogenetic activation of oocytes from large follicles produced higher developmental rates than oocytes from large follicles (p < 0.05). At 28 h, the IVM medium with oocytes from large follicles contained significantly more 17ß‐oestradiol (E₂) than the medium with oocytes from small follicles (5.55 vs 3.45 ng/ml, respectively; p < 0.05) and at 46 h, the medium with oocytes from small follicles contained significantly more progesterone (P₄) than the medium with oocytes from large follicles (276.7 vs 108.2 ng/ml, respectively, p < 0.05). Porcine oocytes from large follicles have higher nuclear and cytoplasmic maturation capacities, but the differences did not appear to be cAMP‐mediated. Our findings also suggest that COCs from small follicles undergo more intensive luteinization than COCs from large follicles. The results show that oocytes from follicles with a diameter greater than 4.0 mm are more suitable for in vitro studies.     

Noradrenergic Control of Arginine Vasopressin Release from the Ewe Hypothalamus In Vitro: Sensitivity to Oestradiol

The present study aims at ascertaining the influence of α₁‐adrenoreceptors on arginine vasopressin (AVP) release in vitro and determine whether E₂ modulates the α₁‐adrenoreceptor and AVP interaction. Ten minutes after ewe killing, sagittal midline hypothalamic slices (from the anterior preoptic area to the mediobasal hypothalamus with the median eminence, 2 mm thick, 2 per sheep) were dissected, placed in oxygenated minimum essential media‐α (MEM‐α) at 4°C and within 2 h were singly perifused at 37°C with oxygenated MEM‐α (pH 7.4; flow rate 0.15 ml/min), either with or without E₂ (24 pg/ml). After 4 h equilibration, 10 min fractions were collected for 4 h interposed with 10 min exposure at 60 min to a specific α₁‐adrenoreceptor agonist or antagonist at various doses (0.1–10 mm ). At the end of all perifusions, slices responded to KCl (100 mm ) with AVP efflux (p < 0.05). Release of AVP was enhanced (p < 0.05) by the α₁‐adrenoreceptor agonist (methoxamine 10 mm ; no E₂, n = 7 perifusion chambers: from 14.3 ± 2.7 to 20.9 ± 3.9, with E₂, n = 10: from 10.7 ± 1.2 to 18.4 ± 3.4 pg/ml) or the antagonist (thymoxamine 10 mm ; no E₂, n = 5: from 9.5 ± 3.1 to 30.4 ± 6.0, with E₂, n = 10: from 10.8 ± 0.9 to 39.1 ± 6.3 pg/ml). With the agonist, the response occurred only at 80 min (p < 0.05) both in the presence and absence of E₂. Whereas, after the antagonist, values were higher (p < 0.05) throughout the post‐treatment period (80–170 min) without E₂, but declined by 150 min in the presence of E₂. Furthermore, the response to the α₁‐adrenoreceptor antagonist was greater (p < 0.05; 90–140 min) than the agonist only in the presence of E₂. In conclusion, these results reveal direct α₁‐adrenoreceptor‐mediated control of the hypothalamic AVP neuronal system which is modulated by E₂.     

Selection of appropriate endotracheal tube size using thoracic radiography in Beagle dogs

Objective

To determine the optimal endotracheal tube size in Beagle dogs using thoracic radiography.

Noradrenergic Control of GnRH Release from the Ewe Hypothalamus In Vitro: Sensitivity to Oestradiol

The present study investigates the influence of α₁‐adrenoreceptors in GnRH release in vitro and determines whether oestradiol modulates α₁‐adrenoreceptor‐GnRH interaction. Within 10 min after ewe sacrifice, saggital midline hypothalamic slices were dissected, placed in oxygenated Minimum Essential Media‐α (MEM‐α) at 4°C and within 2 h were singly perifused at 37°C with oxygenated MEM‐α (pH 7.4; flow rate 0.15 ml/min), either with or without oestradiol (24 pg/ml). After 4‐h equilibration, 10‐min fractions were collected for 4 h interposed with a 10‐min exposure at 60 min to specific α₁‐adrenoreceptor agonist (methoxamine) or antagonist (thymoxamine) at various doses (0.1–10 mm ). The α₁‐adrenoreceptor agonist (10 mm ) increased (p < 0.05) GnRH release at 90 min both in presence and absence of oestradiol. However, in presence of oestradiol, α₁‐adrenoreceptor agonist (10 mm )‐induced GnRH release remained elevated (p < 0.05) for at least 60 min. The bioactivity of the released GnRH was studied using a hypothalamus–pituitary sequential double‐chamber perifusion. Only after exposure of hypothalamic slices to α₁‐adrenoreceptor agonist (10 mm ), did the hypothalamic eluate stimulate LH release from pituitary fragments (n = 9, 7.8 ± 12.3–36.2 ± 21.6 ng/ml) confirming that the α₁‐adrenoreceptor agonist stimulated release of biologically active GnRH. In summary, GnRH release from the hypothalamus is under stimulatory noradrenergic control and this is potentiated in the presence of oestradiol.     

Intrafollicular and systemic serotonin,oestradiol and progesterone concentrations in cycling mares

The hypothesis that a local serotonergic network might also exist in the follicle of mares remains poorly documented, with exception for humans and laboratory species. For this reason, the aim of the present study was to clarify this possibility, investigating intrafollicular serotonin concentrations of the cycling mare at ovulation time. Sixty ovaries collected from 30 clinically healthy mares of slaughterhouse meat production with clinically normal reproductive tracts after slaughtering were evaluated. Blood samples were taken prior to sacrifice. Follicles were classified in three categories in relation to size, as small (20–30 mm), medium (31–40 mm) and large (>41 mm), and the follicular fluid samples were extracted from each follicle. Intrafollicular and systemic serotonin (5‐HT), oestradiol‐17β (E₂) and progesterone (P₄) were determined by means of enzyme‐linked immunosorbent assay and RIA, respectively. Intrafollicular 5‐HT, E₂ and P₄ concentrations were higher than systemic ones (p < .05). 5‐HT concentrations increased in larger compared to medium follicles, without differences compared to small size follicles (p < .05). 5‐HT and E₂ (r = .79) and 5‐HT and P₄ (r = .79; p < .05) were positively correlated. 5‐HT and P₄ concentrations in follicular fluid increased progressively with the increase in follicular size (p < .05). Follicle diameter and E₂ (r = .85) and P₄ (r = .68) were correlated (p < .05). Since serotonin interacts with steroids, its role on steroidogenesis during growth of the dominant follicle may be suggested.     

COMPUTED TOMOGRAPHY OF BALL PYTHONS (Python regius) IN CURLED RECUMBENCY

Anesthesia and tube restraint methods are often required for computed tomography (CT) of snakes due to their natural tendency to curl up. However, these restraint methods may cause animal stress. The aim of this study was to determine whether the CT appearance of the lungs differs for ball pythons in a curled position vs. tube restraint. Whole body CT was performed on ten clinically healthy ball pythons, first in curled and then in straight positions restrained in a tube. Curved multiplanar reformatted (MPR) lung images from curled position scans were compared with standard MPR lung images from straight position scans. Lung attenuation and thickness were measured at three locations for each scan. Time for positioning and scanning was 12 ± 5 min shorter for curled snakes compared to tube restraint. Lung parenchyma thickness and attenuation declined from cranial to caudal on both straight and curled position images. Mean lung parenchyma thickness was greater in curled images at locations 1 (P = 0.048) and 3 (P = 0.044). Mean lung parenchyma thickness decreased between location 1 and 2 by 86–87% (straight: curled) and between location 1 and 3 by 51‐50% (straight: curled). Mean lung attenuation at location 1 was significantly greater on curled position images than tube restraint images (P = 0.043). Findings indicated that CT evaluation of the lungs is feasible for ball pythons positioned in curled recumbency if curved MPR is available. However, lung parenchyma thickness and attenuation in some locations may vary from those acquired using tube restraint.