OBJECTIVE:To develop a topical sildenafil hydrogel and evaluate its effect on wound healing in dogs.ANIMALS:6 purpose-bred, sexually intact, adult Beagles.PROCEDURES:Hydrogels containing sildenafil citrate, N-methyl-2-pyrrolidone, propylene glycol, and poloxamer 407 were developed. Four excision wounds were created along the dorsum of the dogs. Each wound was treated for 21 days with a nonadherent bandage (C) or with a hydrogel containing 0% (G), 5% (5S), or 10% (10S) sildenafil. Daily bandage changes with wound imaging were performed. Biopsy specimens were collected 5 times.RESULTS:Hydrogels were homogenous at room temperature and released > 90% of the sildenafil within 8 hours in vitro. Time to first granulation tissue was significantly shorter for the sildenafil groups (mean ± SD, 2.8 ± 0.8 days [5S and 10S]), compared with the control groups (5.2 ± 0.4 days [C] and 6.3 ± 1.4 days [G]). The G wounds had a 10% to 14% lower contraction rate, compared with the C, 5S, and 10S wounds. 5S wounds had a total wound area 0.7 ± 0.3 cm2 larger than 10S wounds. No significant differences were present when C wounds were compared with 5S and 10S wounds for total wound area, contraction, or epithelialization. Histologic acute inflammatory scores were higher for 5S and 10S wounds in the early and late stages of wound healing, with higher reparative scores on day 7. Neovascularization was higher for 10S wounds on day 7 and 14.CLINICAL RELEVANCE:The topical sildenafil hydrogel promoted early granulation tissue, which may be beneficial for secondary wound closure in clinical settings.

Abstract Background Keratomycosis is a relatively common, sight threatening condition in horses, where treatment is often prolonged and costly. Subconjunctival (SCo) injections offer less resistance to drug diffusion than the topical route, resulting in better penetration to the ocular anterior segment. Voriconazole, a second generation triazole antifungal, is effective against common fungal organisms causing keratomycosis. If combined with a thermogel biomaterial, voriconazole can be easily injected in the SCo space to provide sustained drug release. The purpose of this study was to evaluate the drug concentrations in the anterior segment and clinical effects after SCo injections of voriconazole-containing thermogel: poly (DL-lactide-co-glycolide-b-ethylene glycol-b-DL-lactide-co-glycolide) (PLGA-PEG-PLGA) in healthy equine eyes. Results Voriconazole aqueous humor (AH) and tear concentrations were compared between 6 horses, receiving 1% voriconazole applied topically (0.2 mL, q4h) (Vori-Top) or 1.7% voriconazole-thermogel (0.3 mL) injected SCo (Vori-Gel). For the Vori-Gel group, voriconazole concentrations were measured in AH and tears at day 2 and then weekly for 23 days, and at day 2 only for the Vori-Top group. Ocular inflammation was assessed weekly (Vori-Gel) using the modified Hackett-McDonald scoring system. Ocular tissue concentrations of voriconazole following SCo 1.7% voriconazole-thermogel (0.3 mL) injections were evaluated post euthanasia in 6 additional horses at 3 different time points. Three horses received bilateral injections at 2 h (n = 3, right eye (OD)) and 48 h (n = 3, left eye (OS)) prior to euthanasia, and 3 horses were injected unilaterally (OS), 7 days prior to euthanasia. Voriconazole-thermogel was easily injected and well tolerated in all cases, with no major adverse effects. On day 2, drug concentrations in tears were higher in the Vori-Top, but not statistically different from Vori-Gel groups. For the Vori-Gel group, voriconazole was non-quantifiable in the AH at any time point. Total voriconazole concentrations in the cornea were above 0.5 μg/g (the target minimum inhibitory concentration (MIC) for Aspergillus sp.) for up to 48 h; however, concentrations were below this MIC at 7 days post treatment. Conclusions Voriconazole-thermogel was easily and safely administered to horses, and provided 48 h of sustained release of voriconazole into the cornea. This drug delivery system warrants further clinical evaluation.

Abstract Background Prolonged cytotoxic concentrations of cytarabine (CA) are required for maximum cytotoxicity. DepoCyt is a human liposomal cytarabine (LC) product that lasts longer in plasma and CSF compared with free CA (FC). The use of LC has not been evaluated in dogs. Objectives To perform a LC pharmacokinetic (PK) study when administered SC in dogs. Animals Five healthy female beagles. Methods Three‐period, 3‐treatment, nonblinded, randomized, and crossover design, including a pilot study. LC was administered at 50 mg/m2 SC and FC was administered at 25 and 50 mg/m2 SC and IV. Plasma CA concentrations were measured until 240, 72, and 8 hours after SC LC, SC FC, and IV FC administration, respectively. CA plasma concentrations were quantitated by ultra‐high‐performance liquid chromatography with mass spectrometry (MS/MS) detection and concentration‐time profiles were evaluated by noncompartmental analysis. Results Subcutaneous LC administration resulted in a maximum plasma concentration of 26.3 to 59.78 ng/mL, time to reach maximum plasma concentration of 2 hours, area under the concentration‐time curve to last measurable concentration of 669.3 to 1126 h × ng/mL, and plasma bioavailability (%F) of 19.6% to 31.3%. The PK profiles of FC after SC and IV administration differed when compared with LC. Conclusions and Clinical Importance In healthy dogs, SC LC administration at 50 mg/m2 results in measurable plasma CA concentrations, is apparently safe and well tolerated, but does not result in prolonged cytotoxic plasma concentrations. Poor absorption of LC prevented establishment of a complete LC PK profile.

Voriconazole (VRC) is a potential treatment for pneumomycosis in horses. The objectives of this study were to determine if the delivery of Vfend using a Flexineb nebulizer produced clinically significant [VRC] in lower airways. The hypothesis was that [VRC] after delivery by nebulization would be greater in the pulmonary epithelial lining fluid than plasma. A secondary objective was to determine [VRC] in upper airways through the collection of nasopharyngeal wash (NPW) samples. Voriconazole solution [Vfend-6.25 mg/mL, 100 (n = 2), 200 (n = 3), 500 (n = 1) mg] was nebulized once in 6 healthy geldings. Clinical responses, duration of nebulization, and [VRC] at various time points (up to 8 hours) in plasma, bronchoalveolar lavage fluid (BALF) supernatant and cell pellet, and NPW samples were recorded. Voriconazole (Vfend-6.25 mg/mL, 200 mg) was nebulized in 5 additional, healthy geldings, and [VRC] was measured in NPW samples pre- and postnebulization at time points up to 8 hours. The antifungal activity of BALF and NPW samples was determined using agar disk diffusion. Concentrations of voriconazole were below detection in plasma, BALF supernatant, and cell pellets for all time points and doses except the BALF cell pellet (0.4 μg/g) immediately after nebulization of 500 mg. For 5 horses, administered 200 mg of Vfend, mean [VCR] in NPW at the end of nebulization and 1, 6, and 8 hours postnebulization were: 30.8 ± 29, 1.0 ± 0.84, 0.2 ± 0.19, and 0.34 ± 0.67 μg/mL, respectively. Only NPW samples obtained immediately postnebulization showed antifungal activity. A nebulized Vfend solution is not recommended for the treatment of pneumomycosis in horses.

BackgroundThe effects of sacubitril/valsartan (S/V) on the renin‐angiotensin‐aldosterone system (RAAS) in dogs with cardiomegaly secondary to myxomatous mitral valve disease (MMVD) are currently unknown.ObjectivesTo determine the pharmacodynamic effects of S/V on the RAAS, natriuretic peptide concentrations, systolic arterial pressure (SAP), tests of renal function, and serum electrolyte concentrations in dogs with cardiomegaly secondary to MMVD.AnimalsThirteen client‐owned dogs weighing 4‐15 kg with American College of Veterinary Internal Medicine (ACVIM) Stage B2 MMVD.MethodsProspective, randomized, double‐blind, placebo‐controlled pilot study of S/V in dogs with ACVIM Stage B2 MMVD.ResultsThirteen dogs were recruited: S/V (n = 7) and placebo (n = 6). The median percentage increase in urinary aldosterone to creatinine ratio (UAldo : C) between day 0 and day 30 was significantly lower in the S/V group (12%;P = .032) as compared with the placebo group (195%). The median percentage decrease of NT‐proBNP concentration from day 0 to day 30 was not statistically different between groups (P = .68). No statistical differences were seen in echocardiographic, thoracic radiographic, SAP, or serum biochemical test results measured at any time point between groups. No adverse events were observed for dogs in either group.Conclusion and Clinical ImportanceSacubitril/valsartan may provide a new pharmaceutical method to effectively inhibit the RAAS in dogs with ACVIM Stage B2 MMVD.

Nepafenac is a water-insoluble nonsteroidal antiinflammatory drug that is available as an ophthalmic suspension (Nevanac®). Suspensions are undesirable for 2 reasons: they tend to cause foreign body sensation and lacrimation, which could limit residence time and drug bioavailability. This decreases the amount of time the drug has to reach the site of action, the cornea. Previously, we improved the solubility and ocular permeability of nepafenac by complexing the drug with hydroxypropyl-β-cyclodextrin. In this study, we used the complex to formulate an ion-activated in situ gel system using sodium alginate, Protanal PH 1033, to increase the residence time and to reduce repeat eye drop instillation. Rheological properties of the formulations revealed that the viscosity of the optimized formulation was increased 30-fold when exposed to the simulated tear fluid (35°C). Permeation studies showed that the drug concentration of the in situ formulations were approximately 10 times higher than the commercial product, Nevanac® (p < 0.001). In addition, the in situ gel formulations had 5-fold higher concentrations of nepafenac retained in the cornea when compared to Nevanac® (p <0.001). Finally, ex vivo drug distribution studies in the porcine eye perfusion model revealed a higher drug retention in various ocular tissues such as cornea, sclera, retina, as compared to Nevanac®.

PurposeTo determine in vitro release profiles, transcorneal permeation, and ocular injection characteristics of a voriconazole-containing thermogel suitable for injection into the subconjunctival space (SCS).MethodsIn vitro release rate of voriconazole (0.3% and 1.5%) from poly (DL-lactide-co-glycolide-b-ethylene glycol-b-DL-lactide-co-glycolide) (PLGA-PEG-PLGA) thermogel was determined for 28 days. A Franz cell diffusion chamber was used to evaluate equine transcorneal and transscleral permeation of voriconazole (1.5% topical solution, 0.3% and 1.5% voriconazole-thermogel) for 24 hours. Antifungal activity of voriconazole released from the 1.5% voriconazole-thermogel was determined via the agar disk diffusion method. Ex vivo equine eyes were injected with liquid voriconazole-thermogel (4°C). Distension of the SCS was assessed ultrasonographically and macroscopically. SCS voriconazole-thermogel injections were performed in a horse 1 week and 2 hours before euthanasia and histopathologic analysis of ocular tissues performed.ResultsVoriconazole was released from the PLGA-PEG-PLGA thermogel for more than 21 days in all groups. Release followed first-order kinetics. Voriconazole diffused through the cornea and sclera in all groups. Permeation was greater through the sclerae than corneas. Voriconazole released from the 1.5% voriconazole-thermogel showed antifungal activity in vitro. Voriconazole-thermogel was easily able to be injected into the dorsal SCS where it formed a discrete gel deposit. Voriconazole-thermogel was easily injected in vivo and did not induce any adverse reactions.ConclusionsVoriconazole-containing thermogels have potential application in treatment of keratomycosis. Further research is required to evaluate their performance in vivo.

Thirty dairy bull calves with normal ophthalmic examinations and culture-negative for Moraxella bovis were randomly assigned to 3 groups for a single eye block, randomized, blinded challenge study. Calves were housed in an accredited isolation facility. In calves assigned to Groups 1 and 2, a 0.6 cm corneal lesion was made on the left central cornea utilizing n-heptanol, and immediately inoculated topically with 1.0 x 107 of M. bovis (strain Epp63-300; origin: NADC). The left eye of calves in Group 3 was inoculated topically with M. bovis, but nothing further. In Group 1,2 mL of 0.009% hypochlorous acid (Vetericyn Plus™ Pinkeye Spray) was sprayed topically onto each calf's cornea twice daily for 10 days. In Group 2, 2 mL of 0.9% saline was administered topically to each calf’s cornea twice daily for 10 days. Each animal was scored for ocular pain twice daily. Samples from all eyes were collected for culture on days -7, 0,1,4, and 10. Eyes of all calves were stained daily with fluorescein, and digital photographs were taken of the lesion to assess healing of the cornea. Additionally, serum and plasma samples were collected from all calves on days 0,1,10,11, and 17, and changes in sodium and chloride levels were evaluated. Urine samples and liver, muscle, and fat biopsies were collected from all calves on days 0,11, and 17 and evaluated for chlorine utilizing the N,N-diethyl- p-phenylenediamine (DPD) colorimetric method. Only calves in Group 2 were culture-positive for M. bovis during the study period. The average pain score and days to cure for Group 1 was lower than Group 2 (P < 0.05 and P < 0.02, respectively). Corneal lesion circumference of calves in Groups 1 and Group 2 differed (P < 0.04), and the difference between days was significantly different (P < 0.0001). There was no difference in Na or Cl levels in the plasma and serum samples among the 3 treatment groups at any sampling time points (P< 0.0001), and no difference between pre- (day 0) and post-treatment (day 11 and 17) in chlorine levels in muscle, liver, fat, and urine at any time point. Results of this study suggest that hypochlorous acid spray can be utilized as alternative therapy to reduce pain, infection, and healing time of corneal lesions in calves experimentally infected with M. bovis.

Objective —To develop a high-speed, continuous-flow, automated plasmapheresis procedure for the high-volume harvest of equine plasma in accordance with current good manufacturing practice. Animals —143 horses (predominantly draft breeds) between 3 and 10 years of age at the time of purchase. Procedures —Adaptations were made to automated plasmapheresis instruments and sterile disposable collection sets, which allowed for dual-instrument, continuous-flow operation. Donor horses were connected to the apparatus via 2 catheters (1 inserted in each jugular vein). The instruments removed whole blood from donors, fractionated the blood, diverted plasma to collection bags, and simultaneously returned concentrated cells to the donors. Plasmapheresis was performed on donor horses at 14-day intervals with a maximum of 22 mL of plasma/kg of donor body weight harvested during each plasmapheresis procedure. Results —During a 5-year period, 3,240 plasmapheresis procedures were performed and > 50,000 L of sterile equine plasma was harvested in accordance with current good manufacturing practice. Donors typically remained calm during the plasmapheresis procedures and tolerated the procedures well. The high-volume and frequent plasma harvest did not result in sustained hypoproteinemia in donor horses. Adverse events associated with the automated plasmapheresis technique were infrequent, and the recurrence of adverse events was minimized by making minor adjustments to the procedure. Conclusions and Clinical Relevance —The automated plasmapheresis procedure described in this report can be used to safely harvest equine plasma or to perform therapeutic plasmapheresis in horses.

OBJECTIVE:To determine the effects of intensive serial plasmapheresis on total plasma protein and total IgG concentrations in donor horses involved in a plasmapheresis program.ANIMALS:18 horses (13 mares and 5 geldings; 13 Belgians, 3 Percherons, 1 Standardbred, and 1 warmblood) ranging from 7 to 14 years of age (mean ± SD, 10 ± 3 years) and weighing 822 ± 128 kg.PROCEDURES:Horses from which 22 mL of plasma/kg of donor body weight was harvested at 14-day intervals for a minimum of 8 consecutive plasmapheresis donations were retrospectively selected for use in the evaluation. Automated plasmapheresis procedures were performed by use of 2 modified plasmapheresis instruments/donor horse. Plasma samples were obtained at each donation and used for determination of total protein and total IgG concentrations. Total plasma protein concentrations were determined via refractometry. A commercially available ELISA was used to determine total equine IgG concentrations.RESULTS:The 18 donor horses were used in 8 to 19 serial donations (mean ± SD, 13 ± 3 donations) during the study. Donor horses had significant decreases in both plasma protein and IgG concentrations over the study period.CONCLUSIONS AND CLINICAL RELEVANCE:Serial plasmapheresis procedures caused significant decreases in both plasma protein and IgG concentrations in donor horses; however, decreases were not physiologically relevant. Performing plasmapheresis in horses in accordance with the evaluated automated plasmapheresis procedures did not result in a critical decrease in total plasma protein or total IgG concentrations.

OBJECTIVETo describe in vitro elution characteristics of amikacin and vancomycin from calcium sulfate hemihydrate 98% (plaster of Paris, POP) beads and characterize eluent inhibition of Staphylococcus spp.STUDY DESIGNExperimental study.METHODSPOP beads were impregnated with amikacin or vancomycin alone or in combination and then incubated alone or in combination for 84 days at 37 degrees C in plastic tubes containing sterile phosphate-buffered saline (PBS). Beads containing no antimicrobial served as negative control. Beads were intermittently moved to a new tube containing drug-free PBS. Antimicrobial was measured in the eluent using a polarized fluorescent immunoassay. Eluent inhibition of Staphylococcus spp. was determined at each time point.RESULTSAntimicrobial release from beads was characterized by an initial rapid phase then a slower phase. Although antimicrobial release from beads occurred throughout the 84 days, most was in the first 24 hours, except for vancomycin alone. Duration of eluent inhibition of Staphylococcus spp. growth ranged from 0.5 (amikacin alone) to 56 days (vancomycin alone). Control eluent did not inhibit bacterial growth.CONCLUSIONSAmikacin elution from POP beads was rapid, inhibiting growth for <24 hours with or without vancomycin. Vancomycin elution was slower and inhibited growth for 56 days alone or for 5 days with amikacin.CLINICAL RELEVANCEVancomycin-impregnated beads appear to be reasonable as a therapeutic option whereas amikacin-impregnated POP beads and amikacin and vancomycin combinations may require further study before considering as a therapeutic option.

OBJECTIVE:To evaluate the pharmacokinetic-pharmacodynamic parameters of enrofloxacin and a low dose of amikacin administered via regional IV limb perfusion (RILP) in standing horses.ANIMALS:14 adult horses.PROCEDURES:Standing horses (7 horses/group) received either enrofloxacin (1.5 mg/kg) or amikacin (250 mg) via RILP (involving tourniquet application) in 1 forelimb. Samples of interstitial fluid (collected via implanted capillary ultrafiltration devices) from the bone marrow (BMIF) of the third metacarpal bone and overlying subcutaneous tissues (STIF), blood, and synovial fluid of the radiocarpal joint were collected prior to (time 0) and at intervals after tourniquet release for determination of drug concentrations. For pharmacokinetic-pharmacodynamic analyses, minimum inhibitory concentrations (MICs) of 16 microg/mL (amikacin) and 0.5 microg/mL (enrofloxacin) were applied.RESULTS:After RILP with enrofloxacin, 3 horses developed vasculitis. The highest synovial fluid concentrations of enrofloxacin and amikacin were detected at time 0; median values (range) were 13.22 microg/mL (0.254 to 167.9 microg/mL) and 26.2 microg/mL (5.78 to 50.0 microg/mL), respectively. Enrofloxacin concentrations exceeded MIC for approximately 24 hours in STIF and synovial fluid and for 36 hours in BMIF. After perfusion of amikacin, concentrations greater than the MIC were not detected in any samples. Effective therapeutic concentrations of enrofloxacin were attained in all samples.CONCLUSIONS AND CLINICAL RELEVANCE:In horses with orthopedic infections, RILP of enrofloxacin (1.5 mg/kg) should be considered as a treatment option. However, care must be taken during administration. A dose of amikacin > 250 mg is recommended to attain effective tissue concentrations via RILP in standing horses.

Ketamine is a dissociative anesthetic commonly used for short-term and induction of anesthesia in bovine species. Ketamine has recently become popular for use as a constant-rate infusion for continuous pain relief. Lidocaine, a local anesthetic, is frequently used for local infiltration and epidural nerve blocks. Two of the most common local blocks in adult cattle are the inverted-L local infiltration and caudal epidural nerve blocks. These blocks are utilized for numerous standing surgical procedures such as cesarean sections, obstetric manipulations, correction of a displaced abomasum and perineal surgery. These animals may occasionally be sent for slaughter following a surgical procedure, and the Food and Drug Administration (FDA) has a zero-tolerance policy for drug residues in meat and milk. There is limited information and no published data on meat and milk withdrawal times for ketamine anesthesia or lidocaine administration via these two different techniques. The objectives of this study were to investigate the pharmacokinetics of ketamine following administration of a single intravenous dose and lidocaine administered via inverted-Land caudal epidural nerve blocks, and to establish the withdrawal times for meat and milk of these two anesthetics in mature Holstein cows.