Comparison of Optisol-GS and neomycin-polymyxin B-gramicidin ophthalmic solution for corneal storage in the dog

The objective of the research was to compare the efficacy of Optisol-GS (OGS, Bausch & Lomb Surgical, Irvine, CA, USA) with triple antibiotic ophthalmic solution (neomycin-polymyxin B-gramicidin, NPG; Bausch & Lomb, Tampa, FL, USA) in preserving the viability of corneal endothelial cells. The study subjects were thirty young to middle-aged dogs with no gross corneal pathology that had been euthanized by pentobarbital overdose for reasons unrelated to this project. Corneal tissues were harvested, analyzed, and randomly assigned to treatment groups: one of two media (OGS or NPG), and one of five storage times (1, 7, 14, 21, or 35 days). Six corneas were stored in each medium for each time period. Corneal endothelial cell viability was evaluated pre- and poststorage by vital staining (trypan blue and alizarin red S), and endothelial cell morphology was evaluated with scanning electron microscopy. Storage in NPG caused significant loss (100%) of endothelial cells after all storage times. OGS storage maintained a high level of endothelial cell viability up to 21 days (98.9% ± 1.3% viability). A significant decrease in percentage viability was also found for OGS-stored corneas between 21 and 35 days, when endothelial cell viability decreased to 61.4% ± 45.9%. The conclusions are that NPG storage at −20 °C is a very poor choice of media for corneal tissue banking if graft clarity is the goal. Storage in Optisol-GS at 4 °C for up to 21 days resulted in significantly higher percentages of viable endothelial cells. Optisol-GS storage should facilitate corneal preservation for canine keratoplasty patients.     

Tolerance of the rabbit cornea to an n-butyl-ester cyanoacrylate adhesive (Vetbond)

Objective An n‐butyl‐ester cyanoacrylate adhesive available for veterinary surgery (Vetbond^®, 3M) was tested in rabbits for corneal irritation. Procedures Two experimental procedures were used on 24 rabbits: injection of the adhesive into an intralamellar corneal pocket (n = 10) and application of the glue to a mid‐stromal corneal defect (n = 14). In both experiments the eyes were examined for 20 days for evidence of corneal irritation and tolerance. At the end of each experiment, histopathologic studies were performed on all corneas. Results The corneal reaction to the intrastromally injected cyanoacrylate was characterized clinically by slight edema and vascularization localized to the vicinity of the adhesive. A moderate foreign body‐type reaction was found histologically. Following application of the adhesive to a central stromal defect, the treated corneas remained totally clear and histopathologic examination showed that the healing process was not altered compared to the controls. The mean retention time of the glue patch was 14 days. Conclusions Intrastromal injection and surface application to a corneal defect of n‐butyl‐ester cyanoacrylate to a corneal defect induced only a mild inflammatory response and did not interfere with the reparative process. These findings suggest that this surgical adhesive would be acceptable for treating corneal ulcerations in animals.     

Effects of selective α2‐adrenergic receptor agonists on electrical field‐stimulated contractions of isolated bronchi in horses

We investigated the effects of different selective α₂‐adrenergic receptor (AR ) agonists (detomidine, medetomidine, xylazine, and brimonidine) on the contractions of horse‐isolated bronchi induced by electrical field stimulation (EFS ) and by carbachol. No effects were observed on the contraction induced by carbachol, while α₂‐AR agonists reduced EFS ‐evoked contractions in a concentration‐related fashion. The rank order of potency (pD ₂) was brimonidine (7.40 ± 0.20) >medetomidine (7.09 ± 0.24) >detomidine (6.13 ± 0.55) >xylazine (4.59 ± 0.16). The maximal effects (E_max) were ?56.3% ± 6.3%, ?40.4% ± 6.9%, ?48.6% ± 9.9%, and ?72.7% ± 12.7% for brimonidine, medetomidine, detomidine, and xylazine, respectively. Adrenergic block by guanethidine enhanced the potency (8.10 ± 0.05, 7.30 ± 0.15, 6.83 ± 0.41, and 5.40 ± 0.22) and the efficacy (?95.2% ± 0.7%, ?45.2% ± 11.7%, ?58.5% ± 9.8%, and ?97.9% ± 0.6%) of brimonidine, medetomidine, detomidine, and xylazine, respectively. Selective α₂‐AR antagonist, atipamezole, competitively antagonized the inhibition of EFS ‐evoked contractions induced by all agonists except xylazine. These results suggest the existence of presynaptic α₂‐AR s on cholinergic neurons, negatively regulating the release of acetylcholine in horse bronchial muscle, and that α₂‐AR agonists may be beneficial against vagally mediated bronchoconstriction.     

Assessment of the usage of biodegradable polymeric matrix in vaginal devices to sustain progesterone release in cows

The usage of timed artificial insemination (TAI) at a low cost leading to better reproductive rates has been the aim of several research groups in the field. Usually during TAI protocols, sustained progesterone (P₄) release devices are employed. Most devices are constituted of a nylon skeleton covered with a silicon layer with P₄. A device based on biopolymers was developed in order to reduce costs and decrease its environmental impact. In this study, we compared the kinetics of sustained progesterone release among devices manufactured with a polymeric blend made of polyhydroxybutyrate‐valerate (PHBV) and poly‐ε‐caprolactone (PCL) (DISP) which were compared with DIB® (Internal Bovine Device) used as the control. In the in vitro and in vivo progesterone release tests, two types of biopolymer‐based devices with a superficial area of 147 cm² were used: DISP8 (46% PHBV, 46% PCL and 8% P₄; n  = 4), DISP10 (45% PHBV, 45% PCL, 10% P₄; n  = 4) and DIB® (1 g P₄, 120 cm² area; n  = 3). The in vitro tests were carried out according to USP XXIII specifications and were performed in a dissolutor sink using an alcohol/water mixture (60/40 v/v) as a release media and samples were collected at 2 min, 2, 4, 8, 12, 24, 48, 60, 72, 84 and 96 h. P₄ concentrations were measured through spectrophotometry in a 244 nm long wave. Three to 3 comparisons of angular coefficients of the straight lines obtained by regression analysis of accumulated P₄ concentrations as a function of square root of time were carried out. Furthermore, the diffusion coefficient values of P₄ were also determined for DISP8 and DISP10. The results showed that the concentrations of P₄ were higher in the DISP10 (774.63 ± 45.26 μg/cm²/t^1/2) compared to DISP8 (566.17 ± 3.68 μg/cm²/t^1/2) (P  < 0.05). However, both DISP10 and DISP8 P₄ concentrations did not differ from DIB® (677.39 ± 16.13 μg/cm²/t^1/2). For the analysis of released quantities per day of the in vitro test, four periods were considered: 0–24, 24–48, 48–72 and 72–96 h. In the first 24 h, DISP8 released significantly less P₄ than DISP10 or DIB®, which did not differ among them. Between 24 and 48 h, DISP10 released significantly more P₄ than DIB®. DISP8 released an intermediate P₄ amount and did not differ significantly from DIB® or DISP10. Between 48 and 72 h, P₄ quantity released by DISP10 was significant higher (P  < 0.01) than that of DIB® and DISP8, which did not differ among themselves. Between 72 and 96 h, DISP10 released significantly more P₄ than DIB®, and DISP8 released an intermediate amount which did not differ from DIB® or DISP10 (P  < 0.01). There was interaction between treatment and time (P  = 0.0024). The diffusion coefficient values were: 1.36 × 10^?8 (cm²/s) for DISP10 and 1.12 × 10^?8 (cm²/s) for DISP8. For the in vivo test, ovariectomized crossbred cows received DIB® (n  = 4) or DISP8 (n  = 8) in an alternate design with a non‐balanced sequence (cross‐over) added of measures repeated in time referring to 16 days of blood samples collection. Samples were analyzed through radioimmunoassay in solid phase using the commercial kit of DPC (Diagnostics Products Corporation). Plasma concentrations of P₄ peaked at 4 h after the placement of the device, this being the only time in which plasma P₄ concentrations differed between DIB® (11.45 ± 1.96) compared with DISP8 (9.23 ± 1.15 ng/mL) (P  = 0.027). On day 8, plasma P₄ concentrations were similar for DIB® (2.44 ± 0.09) and DISP8 (1.89 ± 0.13 ng/mL) (P  = 0.58) showing that both devices were able to keep P₄ concentrations above 1 ng/mL in the plasma of the cow during the 16 day in vivo test. In conclusion, devices manufactured with the blend of PHBV/PCL biopolymers can sustain the release of P₄ in a similar manner as silicon.     

Ophthalmic examination findings in a colony of Screech owls (Megascops asio)

Objective To report ophthalmic findings in the Screech owl (Megascops asio). Sample population Twenty‐three, apparently healthy adult captive Screech owls in Maryland. Procedures OU of all owls underwent complete ophthalmic examination. One randomly assigned eye of each bird was measured by phenol red thread tear test (PRT), and the other eye by Schirmer tear test (STT). TonoVet^® rebound tonometry and TonoPen‐XL^® applanation tonometry were performed in each eye to measure IOP. Conjunctival swabs were cultured from one eye of 10 birds, corneal diameter was measured in OU of eight birds, and streak retinoscopy was performed on OU of seven birds. Ten birds were anesthetized, and A‐scan ultrasonography using a 15‐MHz probe was performed to obtain axial intraocular measurements. Results Ophthalmic abnormalities were noted in 24/46 (52%) of eyes. Median STT result was ≤ 2 mm/min, ranging ≤ 2–6 mm/min, and mean ± SD PRT was 15 ± 4.3 mm/15 s. Mean ± SD IOP were 9 ± 1.8 mmHg TonoVet^®‐P, 14 ± 2.4 mmHg TonoVet^®‐D, and 11 ± 1.9 mmHg TonoPen‐XL^®. Coagulase negative staphylococcal organisms were cultured from all conjunctival swabs. Mean ± SD corneal dimensions were 14.5 ± 0.5 mm vertically and 15.25 ± 0.5 mm horizontally. All refracted birds were within one diopter of emmetropia. Mean ± SD axial distance from the cornea to the anterior lens capsule was 4.03 ± 0.3 mm, from cornea to the posterior lens capsule was 10.8 ± 0.5 mm, and from cornea to sclera was 20.33 ± 0.6 mm. Conclusions This study reports ophthalmic examination findings in Screech owls, and provide means and ranges for various ocular measurements. This is the first report of rebound tonometry and PRT in owls.     

Intraocular pressure,specular microscopy,and prostaglandin E2 concentration in dogs with mature and hypermature cataract

This study aimed to evaluate and correlate intraocular pressure (IOP), endothelial cell density (CD), and hexagonality (HEX), and the aqueous humor prostaglandin E₂ (PGE₂) concentration in dogs with mature (MG, n = 8) and hypermature (HG, n = 8) cataracts. Eight laboratory beagles with no ocular abnormalities were included as a control group (CG). The IOP was measured using a digital applanation tonometer. Noncontact specular microscopy was used to evaluate CD and HEX. Samples of aqueous humor were used to determine prostaglandin E₂ concentration using enzyme‐linked immunoassay. Data were compared by anova and Bonferroni's multiple comparison test, and possible correlations among the PGE₂ aqueous concentration and corneal endothelium cell parameters were assessed by Person′s test (P < 0.05). Average values of IOP (P = 0.45) and CD (P = 0.39) were not significantly different between MG, HM, and CG. Average values of HEX were lower, and PGE₂ concentration was increased in the MG and HG in comparison with CG (P < 0.05); however, such parameters did not change significantly between MG and HG (P > 0.05). PGE₂ values did not correlate with IOP, CD, and HEX in any group (P > 0.05). Although there were a small number of dogs studied, our results demonstrated that cataract progression from mature to hypermature did not have a significant change in PGE2 aqueous concentration, IOP, corneal endothelial cell count, or morphology. In addition, PGE₂ concentration was not correlated with parameters of the corneal endothelium or IOP in dogs with mature or hypermature cataracts.     

Distribution of flunixin meglumine and firocoxib into aqueous humor of horses

Background: Nonsteroidal anti‐inflammatory drugs (NSAIDs) are commonly used systemically for the treatment of inflammatory ocular disease in horses. However, little information exists regarding the ocular penetration of this class of drugs in the horse. Objective: To determine the distribution of orally administered flunixin meglumine and firocoxib into the aqueous humor of horses. Animals: Fifteen healthy adult horses with no evidence of ophthalmic disease. Methods: Horses were randomly assigned to a control group and 2 treatment groups of equal sizes (n = 5). Horses assigned to the treatment groups received an NSAID (flunixin meglumine, 1.1 mg/kg PO q24h or firocoxib, 0.1 mg/kg PO q24h for 7 days). Horses in the control group received no medications. Concentrations of flunixin meglumine and firocoxib in serum and aqueous humor and prostaglandin (PG) E₂ in aqueous humor were determined on days 1, 3, and 5 and aqueous : serum ratios were calculated. Results: Firocoxib penetrated the aqueous humor to a significantly greater extent than did flunixin meglumine at days 3 and 5. Aqueous : serum ratios were 3.59 ± 3.32 and 11.99 ± 4.62% for flunixin meglumine and firocoxib, respectively. Ocular PGE₂ concentrations showed no differences at any time point among study groups. Conclusions and Clinical Importance: Both flunixin meglumine and firocoxib penetrated into the aqueous humor of horses. This study suggests that orally administered firocoxib penetrates the aqueous humor better than orally administered flunixin meglumine at label dosages in the absence of ocular inflammation. Firocoxib should be considered for the treatment of inflammatory ophthalmic lesions in horses at risk for the development of adverse effects associated with nonselective NSAID administration.     

Ophthalmic examination findings in adult pygmy goats (Capra hicus)

Objective To document normal ophthalmic findings and ocular abnormalities in captive adult pygmy goats. Animals studied Ten healthy adult pygmy goats (five male, five female; 5–11 years of age; 26–45 kg body mass) underwent complete ophthalmic examinations. Procedure Direct illumination, diffuse and slit‐beam biomicroscopy, indirect ophthalmoscopy, IOP measurements and Schirmer tear tests were performed. TonoVet^® rebound tonometry, followed by topical application of 0.5% ophthalmic proparacaine, and Tono‐Pen XL^® applanation tonometry were performed in each eye to obtain estimates of IOP. Results Ophthalmic abnormalities included corneal scars and pigmentation, incipient cataracts, lenticular sclerosis, and vitreal veiling. Mean STT values were 15.8 mm/min, with a range of 10–30 mm/min. Mean IOP values were 11.8 mmHg for TonoVet^®‐D, with a range of 9–14 mmHg; 7.9 mmHg for TonoVet^®‐P, with a range of 6–12 mmHg; and 10.8 mmHg for Tono‐Pen XL^®, with a range of 8–14 mmHg. Conclusions Ophthalmic examination findings in adult pygmy goats, including normal means and ranges for STT and IOP measurements, using applanation and rebound tonometry, are provided.     

Measurements of canine aqueous humor inflammatory mediators and the effect of carprofen following anterior chamber paracentesis

Objectives Phase I: To evaluate levels of prostaglandin E₂ (PGE₂), nitrites and nitrates (NO_x), tumor necrosis factor‐alpha (TNF‐α) and expression of inducible cyclo‐oxygenase (COX‐2), nitric oxide synthase (NOS‐2), and matrix metalloproteinases (MMP‐3 and ‐9) in canine aqueous humor following repeated anterior chamber paracenteses (ACP). Phase II: to evaluate the effect of carprofen on PGE₂, NO_x, and TNF‐α in canine aqueous humor following ACP. Animals studied Four beagles in phase I and 8 beagles in phase II. Procedures Phase I: ACP was performed at time (T) 0, 4 and 8 h. Phase II: A randomized, placebo‐controlled cross‐over design with four dogs per group where carprofen was given 4.4 mg/kg/day on day (D) 1, 2 and 3. ACP was performed at T0 and T1.5 on D3. Statistical analysis was performed with repeated measures anova and post hoc Tukey‐Kramer multiple‐comparison procedure. In phase II, TNF‐α level was analyzed with a Wilcoxon signed‐rank test. Results Phase I: PGE₂ significantly increased (P < 0.0001) to plateau at T4. NO_X was decreased at T4 (P < 0.06), but increased at T8 (P < 0.0001). COX‐2 showed detectable expression only at T8. TNF‐α, NOS‐2, MMP‐3 and ‐9 were undetectable at all time points. Phase II: At T1.5, PGE₂ was significantly elevated in both groups but was lower in the carprofen group (P = 0.037). NO_x and TNF‐α did not statistically increase in either group. Conclusions Following ACP, significant increases in PGE₂ levels confirmed inflammation characterized by a rise of COX‐2. The NO_x pathway took longer to induce as compared with PGE₂. Carprofen decreased PGE₂ levels and could help control intraocular inflammation.     

The effect of a single dose of topical 0.005% latanoprost and 2% dorzolamide/0.5% timolol combination on the blood-aqueous barrier in dogs: a pilot study

Objective To determine the effects of 0.005% latanoprost and 2% dorzolamide/0.5% timolol on the blood‐aqueous barrier (BAB) in normal dogs. Animals studied Eight mixed‐breed and pure‐breed dogs. Procedures Baseline anterior chamber fluorophotometry was performed on eight normal dogs. Sodium fluorescein was injected and the dogs were scanned 60–90 min post‐injection. Seventy‐two hours following the baseline scan, one eye received one drop of latanoprost. Fluorophotometry was repeated 4 h after drug administration. Following a washout period, the identical procedure was performed 4 h after the administration of dorzolamide/timolol. The degree of BAB breakdown was determined by comparing the concentrations of fluorescein within the anterior chamber before and after drug administration. BAB breakdown was expressed as a percentage increase in the post‐treatment fluorescein concentration over the baseline concentration: %INC [Fl] = {([Fl]_post – [Fl]_baseline)/[Fl]_baseline} × 100. The percentage increase in fluorescein concentration in the treated eye was compared to that in the nontreated eye using a paired t‐test with significance set at P ≤ 0.05. Results Following administration of latanoprost, the fluorescein in the treated eyes increased 49% (± 58%) from baseline compared to 10% (± 31%) in the untreated eyes (P = 0.016). Following administration of dorzolamide/timolol, the fluorescein concentration increased 38% (± 54%) compared to baseline vs. 24% (± 38%) in the untreated eyes (P = 0.22). Conclusions The results of this study show that topical latanoprost may cause BAB disruption in normal dogs while topical dorzolamide/timolol may have no effect on the BAB in normal dogs.     

Association of adrenocorticotrophin and cortisol concentrations with peripheral blood leukocyte cytokine gene expression in septic and nonseptic neonatal foals

Background

The hypothalamic‐pituitary‐adrenal (HPA) is influenced by the proinflammatory cytokines IL‐6, IL‐1β, and TNF‐α in critically ill humans. Information about the association of cytokines with the HPA axis in neonatal foals is lacking.

Hypothesis/Objectives

The objectives were to describe for hospitalized septic and nonseptic foals (1) temporal changes in blood concentrations of ACTH, and cortisol, and leukocyte cytokine gene expression, and (2) coassociation of these HPA axis hormones with blood leukocyte cytokine gene expression.

Animals

Hospitalized septic foals (N = 15) and hospitalized nonseptic foals (N = 11).

Methods

Blood samples, obtained from study foals at admission (T = 0), and 24 (T = 1), 48 (T = 2), 72 (T = 3), and 96 (T = 4) hours after admission, were processed to isolate RNA from leukocytes and to harvest plasma and serum for hormone assays. Plasma ACTH and serum cortisol concentrations were determined by radioimmunoassay. Leukocyte mRNA expression of IL‐1β IL‐6, IL‐8, IL‐10, and TNF‐α was determined using RT‐PCR.

Results

Cortisol concentrations were greater (P < .05) in foals at admission than at other time points. The expressions of IL‐8 and IL‐10 mRNA were lower (P < .05) at each time point in septic than in nonseptic foals. Among septic foals, ACTH was positively associated (P = .0026) with IL‐6 mRNA expression.

Conclusions

Sepsis influences secretion of the HPA axis hormones and expression of cytokines in foals. A positive association with the HPA axis and IL‐6 expression was detected. The clinical importance of these findings requires additional study.     

Reference values for selected ophthalmic diagnostic tests of the capuchin monkey (Cebus apella)

Purpose To perform selected ophthalmic diagnostic tests in healthy capuchin monkeys (Cebus apella) with the aim of establishing normal physiological reference values for this species. Methods A total of 15 healthy, capuchin monkeys were used to test most of the parameters in this investigation. Five of the 15 monkeys were used for the evaluation of normal conjunctival flora. Ages varied from 6 to 20 years of age. Selected diagnostic ocular tests were performed including Schirmer tear test (STT), tonometry using an applanation tonometer (Tonopen^®), central corneal thickness (CCT) using an ultrasonic pachymeter (Sonomed, Micropach^®, Model 200P+) and culture of the normal conjunctival bacterial flora. Results and discussion Results for selected ocular diagnostic tests investigated here for the capuchin monkey eye were as follows: IOP: 18.4 ± 3.8 mmHg; STT: 14.9 ± 5.1 mm/min; CCT: 0.46 ± 0.03 mm. No statistically significant differences between ages or genders were found for any of the results. Streptococcus sp. and Corynebacterium sp. were isolated from healthy conjunctival and eyelid margins, suggesting they are normal constituents of the conjunctival flora of the capuchin monkey. The data obtained in this investigation will help veterinary ophthalmologists and laboratory animal medicine specialists to more accurately diagnose ocular diseases in the capuchin monkey. These ophthalmic reference values will be particularly useful to diagnose discrete or unusual pathological changes of the capuchin monkey eye.     

Systemic Concentrations of Endogenous and Exogenous FSH in Anoestrous Ewes Superstimulated with Folltropin®-V

The synchronization of follicular waves with medroxyprogesterone acetate (MAP) and oestradiol‐17β (E₂‐17β) prior to ovarian superstimilation in anoestrous ewes reduces the variability in superovulatory responses by an unknown mechanism. Follicle stimulating hormone (FSH) is a primary promoter of antral follicular development, but the relevance of circulating FSH concentrations to the superovulation performance in ewes has not been examined. Eighteen anoestrous Rideau Arcott ewes (May–June) were superovulated with Folltropin^®‐V (porcine FSH), with (n = 8; treated ewes) or without (n = 10; control ewes) a single i.m. dose of 350 μg of E₂‐17β, given on the sixth day of a 14‐day treatment with MAP‐releasing intravaginal sponges (60 mg). The superovulatory treatment, begun 6 days after E₂‐17β injection, consisted of six i.m. applications of Folltropin^®‐V given twice daily (at 08:00 and 16:00 h), followed by an i.m. injection of GnRH (50 μg). Blood samples collected every 8 h throughout the 3‐day treatment, were analysed by radioimmunoassays for concentrations of ovine and porcine FSH, using species‐specific standards and primary antibodies. Serum concentrations of oFSH were greater (p < 0.05) in the controls compared to treated ewes at 40, 64 and 72 h and the variability in mean oFSH concentrations was greater (p < 0.05) in control ewes at 40, 48, 64 and 72 h after the 1^st Folltropin^®‐V injection. There were no differences (p > 0.05) between the two groups in serum concentrations of pFSH. Significant correlations were recorded between the number of corpora lutea (CL) and oFSH concentrations at 8 h (r = 0.72, p < 0.05), 16 h (r=0.63, p < 0.05) and 64 h (r = 0.84, p < 0.01) after the 1^st Folltropin^®‐V injection. The total number of recovered embryos was positively correlated to oFSH concentrations at 56 h (r = 0.69, p < 0.05). We concluded that changes in endogenous FSH concentrations during ovarian superstimulation with pFSH might contribute to the variability in superovulatory responses in ewes.     

Silybin inhibits interleukin-1β-induced production of pro-inflammatory mediators in canine hepatocyte cultures

Au, A. Y., Hasenwinkel, J. M., Frondoza, C. G. Silybin inhibits interleukin‐1β‐induced production of pro‐inflammatory mediators in canine hepatocyte cultures. J. vet. Pharmacol. Therap. 34 , 120–129. Hepatocytes are highly susceptible to cytokine stimulation and are fundamental to liver function. We established primary canine hepatocyte cultures to study effects of anti‐inflammatory agents with hepatoprotective properties. Hepatocyte cultures were incubated with control media alone, silybin (SB), or the more bioavailable silybin–phosphatidylcholine complex (SPC), followed by activation with interleukin‐1 beta (IL‐1β; 10 ng/mL). Inflammatory response was measured by prostaglandin E2 (PGE₂), interleukin‐8 (IL‐8), and monocyte chemotactic protein‐1 (MCP‐1) production and also nuclear factor‐kappa B (NF‐κB) translocation. Hepatocyte cultures continued production of the phenotypic marker albumin for more than 7 days in culture. IL‐1β exposure increased PGE₂, IL‐8, and MCP‐1 production, which was paralleled by NF‐κB translocation from the cytoplasm to the nucleus. Pretreatment with SB and SPC significantly inhibited IL‐1β‐induced production of pro‐inflammatory markers and attenuated NF‐κB nuclear translocation. We demonstrate for the first time that primary canine hepatocyte cultures can be maintained in culture without phenotypic loss. The observation that hepatocyte cultures respond to pro‐inflammatory IL‐1β activation indicates hepatocytes as primary cellular targets of extrinsic IL‐1β. The ability of SB and SPC to inhibit hepatocyte culture activation by IL‐1β reinforces the notion of their hepatoprotective effects. Our primary canine hepatocyte culture model facilitates identification of hepatoprotective agents and their mechanism of action.     

Influence of oxytetracycline on carprofen pharmacodynamics and pharmacokinetics in calves

A tissue cage model of inflammation in calves was used to determine the pharmacokinetic and pharmacodynamic properties of individual carprofen enantiomers, following the administration of the racemate. RS(±) carprofen was administered subcutaneously both alone and in combination with intramuscularly administered oxytetracycline in a four‐period crossover study. Oxytetracycline did not influence the pharmacokinetics of R(?) and S(+) carprofen enantiomers, except for a lower maximum concentration (C_max) of S(+) carprofen in serum after co‐administration with oxytetracycline. S(+) enantiomer means for area under the serum concentration–time curve (AUC_0–96h were 136.9 and 128.3 μg·h/mL and means for the terminal half‐life (T_½k₁₀) were = 12.9 and 17.3 h for carprofen alone and in combination with oxytetracycline, respectively. S(+) carprofen AUC_0–96h in both carprofen treatments and T_½k₁₀ for carprofen alone were lower (P < 0.05) than R(?) carprofen values, indicating a small degree of enantioselectivity in the disposition of the enantiomers. Carprofen inhibition of serum thromboxane B₂ ex vivo was small and significant only at a few sampling times, whereas in vivo exudate prostaglandin (PG)E₂ synthesis inhibition was greater and achieved overall significance between 36 and 72 h (P < 0.05). Inhibition of PGE₂ correlated with mean time to achieve maximum concentrations in exudate of 54 and 42 h for both carprofen treatments for R(?) and S(+) enantiomers, respectively. Carprofen reduction of zymosan‐induced intradermal swelling was not statistically significant. These data provide a basis for the rational use of carprofen with oxytetracycline in calves and indicate that no alteration to carprofen dosage is required when the drugs are co‐administered.     

Myocilin protein levels in the aqueous humor of the glaucomas in selected canine breeds

Objective To compare aqueous humor myocilin protein levels in dogs with the primary glaucomas to those with the secondary glaucomas, primary cataracts, and diabetic cataracts. Materials and methods Four groups were selected, based on diagnosis by the attending veterinary ophthalmologists and included: primary glaucoma (primary open‐angle glaucoma (POAG) and primary closed angle glaucoma (PCAG); n = 155); secondary glaucoma (n = 94); primary (presumed inherited) cataract (n = 142), and diabetic cataract (n = 83). A total of 474 samples (187 males, 263 females, 24 unreported) with average ages of 117 months for the males and 101 months for the females were analyzed. Myocilin protein was measured using the Coomassie staining and Western blot methods relative to a myocilin control. Results Differences were seen between nonglaucomatous (cataractous) and glaucomatous dogs with myocilin levels in glaucomatous eyes being many times higher than those in the cataractous dogs. Primary glaucomatous dogs were found to have an aqueous humor myocilin protein level of 17.30 ± 1.03 units. Secondary glaucomas had the highest level of myocilin in the aqueous humor with 19.27 ± 1.41 units. Diabetic cataractous dogs had the lowest levels of myocilin reported with 6.60 ± 0.88 (mean ± SEM) units. Normal (cataractous) dogs had a myocilin level in the aqueous humor of 8.05 ± 0.86 units. Conclusion Aqueous humor protein levels were elevated, relative to the myocilin control, in both the primary and secondary glaucoma groups compared to the cataract and diabetic cataract groups. Like in the Beagle POAG, aqueous humor myocilin protein levels are increased. Further studies are indicated to investigate the exact role of the aqueous humor myocilin protein in the genesis in increased IOP in these primary glaucomatous breeds.     

Normal features of superficial nondesquamating cells of the rabbit corneal epithelium assessed by scanning electron microscopy

Objective To quantitatively assess surface features of corneal epithelial cells with particular emphasis on regional differences in cell size or shape. Animals studied Female New Zealand White rabbits, aged 11–12 weeks. Procedures Animals were exposed to a light : dark cycle of 14 : 10 h for 2 weeks and then the corneas prepared for scanning electron microscopy (SEM) at 1500 h. Images were taken at central location (C), mid‐periphery (MP), and periphery (P) of the cornea. On prints at 5000×, cell–cell borders were marked, and long (L) and short (S) dimensions measured, and the surface ring‐shaped features outlined and counted. Results Across the epithelial surface cells had from 3 to 11 bordering cells (sides), with 5‐sided cells being more common (mean 32.7 ± 11.3%, SD). L dimensions averaged 26.7, 30.9, and 33.9 µm at the C, MP, and P locations, while S dimensions were 18.5, 21.8, and 24.0 µm, respectively. The L : S ratio was1.523, with averages of 1.567, 1.501, and 1.487 at the three locations. Using an averaged cell dimension, cell density was estimated and found to be 7376, 4405, and 3071 cells/mm² at C, MP, and P locations. Almost all cells were decorated with ring‐shaped features (craters), with the number increasing in relation to cell size and were much higher on more peripheral cells. Conclusions The non‐exfoliating corneal epithelial surface is composed of flat polygonal cells often with 5‐sides cells, which are progressively larger towards the peripheral cornea and more decorated with ring‐shaped features.     

Comparative plasma ampicillin levels and bioavailability of five parenteral ampicillin formulations in ruminant calves

Summary

Plasma ampicillin concentrations were determined in an eight‐ways crossover trial involving six ruminant calves, which were treated intravenously (i.v.) with sodium ampicillin at 15.5 mg/kg and intramuscularly (i.m.) with five different ampicillin trihydrate or ampicillin anhydrate formulations at 7.7 mg/kg. The mean plasma concentration‐time curve (C_p)after intravenous ampicillin sodium administration was described biexponentially, as: C_p = 38.8 e ‐0.0268t + 0.45 e ‐0.0058t.

Intramuscular injection, into the lateral neck, of Ampikel‐20^® and Polyflex^® resulted in 100 per cent bioavailabilities within 12 h post injection (p.i.), but the biological half‐lives (t½>) were different, being 2.1 and 3.8 h, respectively. Ampikel‐20^® produced the hïghest peak plasma drug concentrations (mean C _max:4.8 μg ampicillin/ml). After intramuscular injection of Penbritin® the mean bioavailability for the first 12 h p.i. was 63 per cent, the mean t½>, was 5.9 h, and the mean C_max was 1.8 μg/ml. Treatment with Albipen^® and Duphacillin^® resulted in low plasma ampicillin levels, which were maintained for 3 to 6 days p.i., limited bioavailability during the first 12 h p.i., and a mean t½> of 22.2 and 11.9 h, respectively. Plasma concentrations of ampicillin from four hours onwards after i.m. and s.c. administration of Ampikel‐20^® at a dose level of 15.5 mg/ kg were similar.

The duration of potentially therapeutic plasma ampicillin concentrations after administration of each formulation is presented. Pre‐slaughter withdrawal times for diseased calves are suggested for the different formulations studied.     

Conception rates in ewes after AI with ram semen preserved in milk-egg yolk extenders supplemented with glycerol

This study aimed at comparing the effect of ram semen preserved at 5°C on two milk‐based extenders (UHT skim milk or INRA‐96^®, 5% egg yolk) supplemented with 2% glycerol, and the preservation time (24 and 48 h) on conception rates after cervical AI of ewes. In two field trials, 1198 Merino ewes were cervical AI in spontaneous oestrus. In Experiment 1, pooled semen (6 rams) was extended in UHT‐base (fresh, control) or chilled for 24 h in UHT5Y (UHT‐base 5% egg yolk), INRA5Y (INRA‐96^® 5% egg yolk), UHT5Y2G (UHT5Y 2% glycerol) or INRA5Y2G (INRA5Y 2% glycerol). In Experiment 2, AI was performed with pooled semen (7 rams) used fresh (extended in UHT‐base or UHT5Y2G, control groups) or chilled (extended in UHT5Y2G) for 24 or 48 h. Conception rate was determined by ultrasound 40 days after AI. INRA‐96^®– had similar conception as UHT‐preserved semen (56.7 vs 55.4%, p > 0.05). Addition of 2% glycerol did not modify the results (56.8 vs 55.2%, p > 0.05). Fresh semen extended in UHT‐base, and UHT5Y2G yielded similar conception rates (60 vs 64%, p > 0.05). Preservation for 24 or 48 h in UHT5Y2G gave similar results (49 vs 47%; p > 0.05). In conclusion, ram semen chilled for 24 h in UHT‐ or INRA‐96^®‐based extenders yielded similar results, and glycerol addition did not have a detrimental effect. UHT5Y2G might be used to extend ram semen for fresh AI, or to preserve it for 24 or 48 h with acceptable results.