Pharmacokinetics of florfenicol after intravenous and intramuscular administration in New Zealand White rabbits

The pharmacokinetic disposition and bioavailability of florfenicol (FF) were determined after single intravenous (i.v.) and intramuscular (i.m.) administrations of 25 mg/kg b.w. to ten healthy New Zealand White rabbits. Plasma FF concentrations were determined by high-performance liquid chromatography (HPLC). The plasma pharmacokinetic values for FF were best described by a one-compartment open model. The elimination half-life (t_1/2β) was different (p < 0.05) however, the area under curve (AUC) was similar (p > 0.05) after i.v. and i.m. administrations. FF was rapidly eliminated (t_1/2β 1.49 ± 0.23 h), slowly absorbed and high (F, 88.75 ± 0.22%) after i.m. injection. In addition, FF was widely distributed to the body tissues (V_ss 0.98 ± 0.05 L/kg) after i.v. injection. In this study the time that plasma concentration exceeded the concentration of 2 μg/mL was approximately 6 h. For bacteria with MIC of 2 μg/mL, frequent administration at this dose would be needed to maintain the concentration above the MIC. However, it is possible that rabbit pathogens may have MIC values less than 2 μg/mL which would allow for less frequent administration. Further studies are necessary to identify the range of MIC values for rabbit pathogens and to identify the most appropriate PK-PD parameter needed to predict an effective dose.     

Pharmacokinetics and bioavailability of a long-acting formulation of cephalexin after intramuscular administration to cats

The pharmacokinetic profile and bioavailability of a long-acting formulation of cephalexin after intramuscular administration to cats was investigated. Single intravenous (cephalexin lysine salt) and intramuscular (20% cephalexin monohydrate suspension) were administered to five cats at a dose rate of 10 mg/kg. Serum disposition curves were analyzed by noncompartmental approaches. After intravenous administration, volume of distribution (V_z), total body clearance (Cl_t), elimination constant (λ_z), elimination half-life (t_½λ) and mean residence time (MRT) were: 0.33 ± 0.03 L/kg; 0.14 ± 0.02 L/h kg, 0.42 ± 0.05 h⁻¹, 1.68 ± 0.20 h and 2.11 ± 0.25 h, respectively. Peak serum concentration (C_max), time to peak serum concentration (T_max) and bioavailability after intramuscular administration were 15.67 ± 1.95 μg/mL, 2.00 ± 0.61 h and 83.33 ± 8.74%, respectively.     

Determination of oral tramadol pharmacokinetics in horses

The determination of the pharmacokinetic parameters of tramadol in plasma and a better characterization of its metabolites after oral administration to horses is necessary to design dosage regimens to achieve target plasma concentrations that are associated with analgesia. The purpose of this study was to determine the pharmacokinetics and elimination pattern in urine of tramadol and its metabolites after oral administration to horses. Tramadol was administered orally to six horses and its half-life, T_max and C_max in plasma were 10.1, 0.59 h, and 132.7 ng/mL, respectively. The half-life, T_max and C_max for M1 in plasma were 4.0, 0.59 h, and 28.0 ng/mL, respectively. Tramadol and its metabolites were detectable in urine between 1 and 24 h after the administration. In conclusion, the PK data reported in this study provides information for the design of future studies of tramadol in horses.     

Disposition of marbofloxacin in vulture (Gyps fulvus) after intravenous administration of a single dose

The pharmacokinetics properties of marbofloxacin were studied in adult Eurassian Griffon vulture after single-dose intravenous (IV) administration of 2 mg/kg. Drug concentration in plasma was determined by high-performance liquid chromatography and the data obtained were subjected to compartmental and non-compartmental kinetic analysis. Marbofloxacin presented a volume of distribution at steady-state (Vdss) of 1.51 ± 0.22 L and total plasma clearance (Cl) of 0.109 ± 0.023 L/h kg. The permanence of this drug was long in vultures (T_1/2λ = 12.51 ± 2.52 h; MRT_∞ = 13.54 ± 2.29 h). The optimal dose of marbofloxacin estimated is 2.73 mg/kg per day for the treatment of infections in vultures with MIC₉₀ = 0.2 μg/mL.     

Cytotoxicity of Senecio in macrophages is mediated via its induction of oxidative stress

In Arunachal Pradesh and other sub-Himalayan areas of India, accidental consumption of Senecio plants by yaks is often fatal as the plant contains toxic alkaloids like Seneciophylline. The present investigation was undertaken to demonstrate the pro-oxidant effects of an ethanolic extract of Seneciochrysanthemoides (S-EtOH). S-EtOH impaired viability in macrophages, the IC₅₀ being 13.8 ± 1.11 μg/mL. The effect of S-EtOH (1 μg/mL) on generation of reactive oxygen species (ROS) in macrophages was measured by flow cytometry using 2′,7′-dichlorofluorescein diacetate (H₂DCFDA) where it caused a significant increase in the mean fluorescence channel (MFC) from 8.55 ± 0.03 to 47.32 ± 2.25 (p < 0.001). S-EtOH also effected a 3.8-fold increase in extracellular nitric oxide (NO) generation from 4.90 ± 0.72 μM to 18.79 ± 0.32 μM (p < 0.001), a 2.2-fold increase in intracellular NO production, the MFC increasing from 14.95 ± 0.48 to 33.34 ± 1.66 (p < 0.001), and concomitantly depleted non protein thiols as analyzed by flow cytometry using mercury orange, with a reduction in MFC from 632.5 ± 49.44 to 407.4 ± 12.61 (p < 0.01). Additionally, S-EtOH (14 μg/mL, 24 h) caused apoptosis as evident by increased Annexin V binding and terminal deoxynucleotidyl transferase mediated dUTP DNA nick end labeling. Taken together, the cytotoxicity of S-EtOH can be partly attributed to its capacity to inflict oxidative damage via generation of both reactive oxygen and nitrogen species culminating in apoptosis.     

Pharmacokinetics and Effects of Alkalization after Intravenous Administration of Eltenac in Horses

Eltenac (ELT) [4-(2,6-dichlorophenyl)amino-3-thiophene] is a non-steroidal anti-inflammatory drug (NSAID) that was developed for veterinary use in horses and cattle. The pharmacokinetics of ELT was evaluated in horses at 0.5 mg/kg body weight (BW) after single IV injection after 5 days of repeated IV administration and after a single IV injection in horses previously subjected to 250 mg/kg BW of sodium bicarbonate (NaHCO₃) as an alkalization treatment. The aim was to determine whether blood and subsequent urinary alkalization could modify the pharmacokinetics of ELT. Drug quantification was performed with serum and urine using high performance liquid chromatography with UV-visible detection. The results were also integrated with cyclo-oxygenase-inhibition literature data to review the dosage scheme of ELT in horses. After a single intravenous administration, ELT was characterized by rapid distribution (mean t½_λ1 = 0.18 ± 0.07 hour) and a short elimination half-life (mean t½_λ2 = 2.9 ± 0.68 hour). The volume of distribution was small (V_dss = 253.51 ± 47.55 mL/kg), which is likely because of the high percentage of drug protein binding (approximately 97%). The AUC_0-∞ and Cl_B were 6.92 ± 0.84 h*μg/mL and 73.2 ± 10 mL/h/kg, respectively. Repeated administration did not cause either accumulation or modification of the pharmacokinetic profile. The in vitro effective concentrations were maintained for a 6-hour period. The alkalization procedure appeared to accelerate drug elimination, as ELT was quantifiable only for 6 hours; however, the drug clearance was not significantly modified. Thus, the administration of alkaline compounds to accelerate the elimination of ELT is not completely confirmed.     

Regional differences in transdermal penetration of fentanyl through equine skin

The rate and regional differences for the penetration of fentanyl through equine skin was investigated in vitro using a commercial transdermal therapeutic system (TTS) or ‘patch’. Skin collected from the thorax, groin and leg (dorsal metacarpal) regions of five horses was placed in diffusion cells and a fentanyl TTS applied to each skin sample. Drug penetration through each skin sample over 48 h measured using high performance liquid chromatography (HPLC). Cumulative penetration (μg/cm²) was plotted against time (h) and used to regress the steady state flux (μg/cm²/h) of fentanyl through each skin site. Results showed similar fluxes for both the thorax (2.32 ± 0.17 μg/cm²/h and groin (2.21 ± 0.11 (μg/cm²/h) regions, but significantly lower flux (P = < 0.05) for the leg region (1.56 ± 0.120 μg/cm²/h. Interestingly, there was a significantly longer lag time for the penetration of fentanyl through the groin region (7.87 ± 0.51 h) compared to the other two sites (5.66 ± 0.97 h and 5.75 ± 0.43 h for the thorax and leg regions respectively). The results suggest that a fentanyl TTS applied to the leg region may have a small but significantly lower amount of fentanyl available systemically, compared to patches applied to the thorax or groin regions, which may affect the level of analgesia subsequently achieved in the horse.     

Pharmacokinetics and bioavailability of moxifloxacin in buffalo calves

The pharmacokinetics of moxifloxacin were investigated in buffalo calves following a single intravenous and intramuscular administration of moxifloxacin (5 mg kg⁻¹ body wt.). Moxifloxacin concentrations in plasma and urine were determined by microbiological assay. Pharmacokinetic analysis of disposition data indicated that intravenous administration data were best described by a two compartment open model, whereas intramuscular administration data were best described by a one compartment open model. Following intravenous administration, the elimination half life (t_1/2β), volume of distribution (Vd_(area)) and total body clearance were 2.69 ± 0.14 h, 1.43 ± 0.08 L kg⁻¹ and 371.2 ± 11.2 ml kg⁻¹ h⁻¹, respectively. Following intramuscular administration, the absorption half life (t_1/2ka) was 0.83 ± 0.20 h. The systemic bioavailability (F) of moxifloxacin in buffalo calves was 80.0 ± 4.08%. Urinary excretion of moxifloxacin was less than 14% after 24 h of administration of drug. In vitro binding of moxifloxacin to plasma proteins of buffalo calves was 28.4 ± 3.77%. From the data of surrogate markers (AUC/MIC, C_max/MIC), it was determined in the buffalo calves that when administered by intravenous or intramuscular route at 5 mg kg⁻¹, moxifloxacin is likely to be effective against bacterial isolates with MIC ? 0.1 μg ml⁻¹.     

Combined subcutaneous administration of ivermectin and nitroxynil in sheep: Age/body weight related changes to the kinetic disposition of both compounds

The effect of age/body weight in the plasma disposition kinetics of ivermectin (IVM) and nitroxynil (NTX) after their co-administration as a combined formulation to sheep was studied. Sixteen (16) male sheep were allocated into two experimental groups (n = 8 each): (a) high body weight (high bw) (18-20 months old), and (b) low body weight (low bw) (6-8 months old). Animals in both groups were subcutaneously (sc) treated with IVM (200 μg/kg) and NTX (10 mg/kg) using a commercially available combined formulation (Nitromectin^®, Lab. Ovejero, Spain). Blood samples were taken by jugular venopuncture before (time 0), at 2, 4, 8, 12 h and at 1, 2, 3, 5, 7, 10, 15, 20, 25, 35, 40, 50 and 60 days after administration. Recovered plasma was analysed to quantify IVM and NTX by HPLC. Higher IVM plasma concentrations were measured until 20 days post-administration in “low bw” compared to “high bw” animals, where IVM was recovered up to 35 days post-treatment. The IVM absorption process greatly differed between experimental groups. A significantly higher (p < 0.01) C_max (36.7 ± 7.52 ng/ml) value was obtained at a delayed (p < 0.05) T_max (48.0 ± 0.0 h) in light compared to heavy (C_max: 8.0 ± 0.80 ng/ml; at 34.0 h) body weight sheep. IVM elimination half-life and mean residence time were significantly shorter in light compared to heavy (older) sheep. NTX mean plasma concentrations were lower in “low bw” compared to those measured in “high bw” sheep, with elimination phases declining up to 60 d post-administration in both experimental groups. The NTX AUC value in “low bw” (1188.5 ± 122.6 μg day/ml) was significantly lower (p < 0.05) than that obtained in the “high bw” (oldest) animals (1735.0 ± 155.8 μg day/ml). Shorter NTX elimination half-life and mean residence time (p < 0.01) were obtained in the youngest (“low bw”) compared to the oldest (high bw) sheep. The work reported here assessed for the first time the disposition of IVM and NTX after their combinated injection to sheep, demonstrating that animal body weight/development greatly affects the kinetic behaviour of both anthelmintic drugs.     

Purification of cattle oviduct specific proteins and their effect on in vitro embryo development

The purpose of this study was to determine the effect of oviduct specific proteins as a media supplement for in vitro embryo development in cattle. The proteins were extracted from oviducts of cows and precipitated by ammonium sulfate (30%, 40%, 50% and 60%) followed by dialysis in 50 mM Tris–HCl (pH 7.0) buffer. The dialyzed proteins were fractionated into acidic, basic and neutral fractions using SP sephadex cation exchange and DEAE sephadex anion exchange column chromatography respectively. Cow oviduct specific proteins (cOSPs) constituting all the extracted proteins were used as media supplement in three different concentrations (10, 50 and 100 μg/ml) for in vitro maturation, fertilization and culture (IVMFC) of cow oocytes. Acidic, basic and neutral (unbound) fractions were also used as media supplement in three different concentrations (10, 30 and 50 μg/ml) for IVMFC. Cumulus oocytes complexes were collected from slaughterhouse ovaries, washed thoroughly and cultured in maturation media for 24 h in 5% CO₂ at 38.5 °C with maximum humidity. In vitro matured oocytes were co-incubated with in vitro capacitated sperm in Fert-BO media at 38.5 °C for 18 h in 5% CO₂. The fertilized oocytes were washed and cultured in embryo development media for cleavage. After 40–42 h cleavage was observed and embryos were put in the replacement media for further development. The cleavage rates (%) for cOSPs were observed as 68.24±2.46, 69.28±2.05, 61.77±0.93 and 42.62±1.31 at concentrations of 0, 10, 50 and 100 μg/ml respectively. Rates of blastocyst stage development were 14.49±3.61, 21.17±2.77, 14.66±1.06 and 11.98±1.84. These results indicate that addition of cOSP at10 μg/ml increased blastocyst formation as compared to other concentrations (0, 50 and 100 μg/ml). Although acidic, basic and neutral fractions seemed to have no major effect on cleavage rate, but both acidic and neutral fraction of oviduct specific proteins improved the cleavage rate at 30 μg/ml concentration and basic fraction improved the blastocyst formation at 10 μg/ml concentration.     

Insights into the mechanism by which kisspeptin stimulates a preovulatory LH surge and ovulation in seasonally acyclic ewes: Potential role of estradiol

We have previously demonstrated that a constant intravenous infusion of kisspeptin (Kp) for 48 h in anestrous ewes induces a preovulatory luteinizing hormone (LH) surge followed by ovulation in approximately 75% of animals. The mechanisms underlying this effect are unknown. In this study, we investigated whether Kp-induced preovulatory LH surges in anestrous ewes were the result of the general activation of the whole gonadotropic axis or of the direct activation of central GnRH neurons required for the GnRH/LH surge. In the first experiment, a constant iv infusion of ovine kisspeptin 10 (Kp; 15.2 nmol/h) was given to 11 seasonally acyclic ewes over 43 h. Blood samples were taken every 10 min for 15 h, starting 5 h before the infusion, and then hourly until the end of the infusion. We found that the infusion of Kp induced a well-synchronized LH surge (around 22 h after the start of the Kp infusion) in 82% of the animals. In all ewes with an LH surge, there was an immediate but transient increase in the plasma concentrations of LH, follicle-stimulating hormone (FSH), and growth hormone (GH) at the start of the Kp infusion. Mean (± SEM) concentrations for the 5-h periods preceding and following the start of the Kp infusion were, respectively, 0.33 ± 0.09 vs 2.83 ± 0.49 ng/mL (P = 0.004) for LH, 0.43 ± 0.05 vs 0.55 ± 0.03 ng/mL (P = 0.015) for FSH, and 9.34 ± 1.01 vs 11.51 ± 0.92 ng/mL (P = 0.004) for GH. In the first experiment, surges of LH were observed only in ewes that also had a sustained rise in plasma concentrations of estradiol (E₂) in response to Kp. Therefore, a second experiment was undertaken to determine the minimum duration of Kp infusion necessary to induce such a pronounced and prolonged increase in plasma E₂ concentration. Kisspeptin (15.2 nmol/h) was infused for 6, 12, or 24 h in seasonally acyclic ewes (N = 8), and blood samples were collected hourly for 28 h (beginning 5 h before the start of infusion), then every 2 h for the following 22 h. Kisspeptin infused for 24 h induced LH surges in 75% of animals, and this percentage decreased with the duration of the infusion (12 h = 50%; 6 h = 12.5%). The plasma concentration of E₂ was greater in ewes with an LH surge compared to those without LH surges; mean (± SEM) concentrations for the 5-h period following the Kp infusion were, respectively, 2.23 ± 0.16 vs 1.27 ± 0.13 pg/mL (P < 0.001). Collectively, our results strongly suggest that the systemic delivery of Kp induced LH surges by activating E₂-positive feedback on gonadotropin secretion in acyclic ewes.     

Pharmacokinetics and Effects of Alkalization During Oral and Intravenous Administration of Naproxen in Horses

The use of suitable therapeutic protocols is particularly important when extra-label drugs are used or when physiological parameters are modified, as in the case of the administration of alkaline substances to racehorses. The pharmacokinetics of naproxen (NAP), after both intravenous (iv) and oral administration of 10 mg/kg body weight (BW), was investigated in horses under normal metabolic conditions and in horses whose conditions were modified by the iv administration of 250 mg/kg BW of sodium bicarbonate (NaHCO₃). The hypothesis that blood and consequent urinary alkalization could modify NAP pharmacokinetics was evaluated. Drug quantification was performed on serum and urine using High Performance Liquid Chromatography (HPLC) with ultraviolet-visible detection. Results were also integrated with cycloxygenase (COX)-inhibition published data to suggest an appropriate schedule for NAP dosage in horses. After iv administration, NAP was rapidly distributed (t_1/2α: 0.71 ± 0.43 iv NaHCO₃ and 0.55 ± 0.62 hours No NaHCO₃), whereas its elimination was quite slow (t_1/2β: 6.74 ± 0.41 hours), particularly in iv NaHCO₃ animals (t_1/2β: 8.95 ± 1.37 hours). After oral treatments, NAP was more rapidly absorbed and elimination was slower in iv NaHCO₃ animals (t_1/2λ_z: 17.50 ± 6.66 vs. 7.17 ± 0.91 hours). The oral bioavailability of NAP was approximately 87% and 77% in No NaHCO₃ and iv NaHCO₃, respectively. Urinary excretion of the drug as a parent compound was low. The alkalization procedure did not anticipate the elimination of the acidic drug as expected, but it also influenced the absorption of the drug that was administered orally. The dosage scheme of 10 mg/kg BW iv or orally seems to be appropriate to produce an anti-inflammatory effect for 12 to 24 hours.     

Conditioning Horses at v10 3 Times per Week Does Not Enhance v4

This study examined the effect of exercising horses 3 times per week with two bouts of 5-minutes' duration at their v₁₀. Six Thoroughbreds were treadmill-conditioned for 6 weeks. A standardized exercise test (SET) was performed at the beginning of the conditioning period to determine the blood lactate–running speed (BLRS) relation, and the SET was repeated every 2 weeks. After each SET, the BLRS relation was used to calculate the horse's speed, which produced a blood lactate (LA) concentration of 10 mmol/L (v₁₀) and 4 mmol/L (v₄). Each horse was then conditioned for the next 2 weeks (3 times/week) at its individual v₁₀ for two 5-minute bouts with a 5-minute walking phase in between. Exercise speed was individually adapted to the new v₁₀ every 2 weeks. The v₄ of horses decreased after the first 2 weeks (from 6.23 ± 0.41 m/s to 5.95 ± 0.33 m/s, mean ± SD; P < .05), increased in the following 2 weeks (6.33 ± 0.58 m/s; P < .01), and stayed constant thereafter (P > .05). The conclusion drawn was that exercising horses 3 times per week at their v₁₀ for two 5-minute bouts did not improve v₄.     

Haemosiderin deposition in Donkey (Equus asinus) livers: Comparison of quantitative histochemistry for iron and liver iron content

Post mortem liver samples from 12 donkeys (Equus asinus) aged 21-57 years (4 females, 1 stallion, 7 geldings), were assessed chemically for copper and iron content on a wet weight basis and histologically for stainable iron. Chemical liver copper content ranged from 2.7 to 4.8 μg/g (mean 3.5 ± 0.05 μg/g). Chemical liver iron content ranged from 524 to 5010 μg/g (mean 1723 ± 1258 μg/g). Histochemical iron was measured morphometrically using a computer-based image analysis system; percentage section area staining for iron ranged from 0.84% to 26.69% (mean 10.82 ± 8.36%). There was no clear correlation, within the wide range of iron values, between histochemically demonstrable iron and chemically measured iron content. No clear age-related increase was apparent for either parameter in these aged donkeys. The accumulation of iron in the liver of donkeys may represent a physiological haemosiderosis rather than pathological haemochromatosis.     

Effect of gentle stroking and vocalization on behaviour,mucosal immunity and upper respiratory disease in anxious shelter cats

Emotional, behavioural, and health benefits of gentle stroking and vocalizations, otherwise known as gentling, have been documented for several species, but little is known about the effect of gentling on cats in stressful situations. In this study, 139 cats rated as anxious upon admission to an animal shelter were allocated to either a Gentled or Control group. Cats were gentled four times daily for 10 min over a period of 10 days, with the aid of a tool for cats that were too aggressive to handle. The cats’ mood, or persistent emotional state, was rated daily for 10 d as Anxious, Frustrated or Content. Gentled cats were less likely to have negatively valenced moods (Anxious or Frustrated) than Control cats (Incidence Rate Ratio [IRR] = 0.61 CI 0.42–0.88, P = 0.007). Total secretory immunoglobulin A (S-IgA) was quantified from faeces by enzyme-linked immunosorbent assay. Gentled cats had increased S-IgA (6.9 ± 0.7 log_e μg/g) compared to Control cats (5.9 ± 0.5 log_e μg/g) (P < 0.0001). Within the Gentled group of cats, S-IgA values were higher for cats that responded positively to gentling (7.03 ± 0.6, log_e μg/g), compared with those that responded negatively (6.14 ± 0.8, log_e μg/g). Combined conjunctival and oropharyngeal swab specimens were tested by quantitative real-time polymerase chain reaction (rPCR) for feline herpesvirus type 1 (FHV-1), feline calicivirus (FCV), Mycoplasma felis, Chlamydophila felis, and Bordetella bronchiseptica. There was a significant increase in shedding over time in Control cats (23%, 35%, 52% on days 1, 4 and 10, respectively), but not in gentled cats (32%, 26%, 30% on days 1, 4 and 10, respectively) (P = 0.001). Onset of upper respiratory disease was determined by veterinary staff based on clinical signs, in particular ocular and/or nasal discharge. Control cats were 2.4 (CI: 1.35–4.15) times more likely to develop upper respiratory disease over time than gentled cats (P < 0.0001). It is concluded that gentling anxious cats in animal shelters can induce positive affect (contentment), increase production of S-IgA, and reduce the incidence of upper respiratory disease.     

Pharmacokinetics of tobramycin following intravenous,intramuscular, and intra‐articular administration in healthy horses

The objectives of this study were to examine the pharmacokinetics of tobramycin in the horse following intravenous (IV), intramuscular (IM), and intra‐articular (IA) administration. Six mares received 4 mg/kg tobramycin IV, IM, and IV with concurrent IA administration (IV+IA) in a randomized 3‐way crossover design. A washout period of at least 7 days was allotted between experiments. After IV administration, the volume of distribution, clearance, and half‐life were 0.18 ± 0.04 L/kg, 1.18 ± 0.32 mL·kg/min, and 4.61 ± 1.10 h, respectively. Concurrent IA administration could not be demonstrated to influence IV pharmacokinetics. The mean maximum plasma concentration (C_max) after IM administration was 18.24 ± 9.23 μg/mL at 1.0 h (range 1.0–2.0 h), with a mean bioavailability of 81.22 ± 44.05%. Intramuscular administration was well tolerated, despite the high volume of drug administered (50 mL per 500 kg horse). Trough concentrations at 24 h were below 2 μg/mL in all horses after all routes of administration. Specifically, trough concentrations at 24 h were 0.04 ± 0.01 μg/mL for the IV route, 0.04 ± 0.02 μg/mL for the IV/IA route, and 0.02 ± 0.02 for the IM route. An additional six mares received IA administration of 240 mg tobramycin. Synovial fluid concentrations were 3056.47 ± 1310.89 μg/mL at 30 min after administration, and they persisted for up to 48 h with concentrations of 14.80 ± 7.47 μg/mL. Tobramycin IA resulted in a mild chemical synovitis as evidenced by an increase in synovial fluid cell count and total protein, but appeared to be safe for administration. Monte Carlo simulations suggest that tobramycin would be effective against bacteria with a minimum inhibitory concentration (MIC) of 2 μg/mL for IV administration and 1 μg/mL for IM administration based on C_max:MIC of 10.     

A zoospore inhibition technique to evaluate the activity of antifungal compounds against Batrachochytrium dendrobatidis and unsuccessful treatment of experimentally infected green tree frogs (Litoria caerulea) by fluconazole and benzalkonium chloride

Effective and safe treatments of chytridiomycosis in amphibians, caused by Batrachochytrium dendrobatidis, are needed to help prevent mortality in captive programs for threatened species, to reduce the risk of spread, and to better manage the disease in threatened populations. We describe a simple method to determine minimum inhibitory concentrations (MICs) of antifungal agents that involves adding zoospores to various drug concentrations in 96 well plates and microscopic observation after four days. We report results from testing 10 commercially available antifungal compounds: benzalkonium chloride (<0.78 μg/ml), povidone iodine (312.5 μg/ml), amphotericin B (3.125 μg/ml), fluconazole (<1.56 μg/ml), itraconazole (<1.56 μg/ml), enilconazole (<1.56 μg/ml), mercurochrome (6.25 μg/ml), sodium chloride (12.5 mg/ml), methylene blue (<1.56 μg/ml) and Virkon (3.125 μg/ml). For treatment trials of juvenile Litoria caerulea, baths of benzalkonium chloride at 1 mg/L and fluconazole at 25 mg/L were used on 18 experimentally infected frogs per treatment. Although these treatments resulted in longer survival times (mean 43.7 ± 11.3 days) than in the untreated controls (37.9 ± 9.3 days), the mortality rate was still 100%. Higher doses of fluconazole are suggested for further animal trials.     

Effects of mosapride citrate, metoclopramide hydrochloride, lidocaine hydrochloride, and cisapride citrate on equine gastric emptying, small intestinal and caecal motility

Objective

Although extensive work has been done to elucidate the beneficial and unfavorable effects of gastrointestinal prokinetic agents in humans, little is known on the effects of these agents in horses. In this study, we compared the effects of mosapride, metoclopramide, cisapride, and lidocaine on equine gastric emptying, jejunal and caecal motility and evaluated these agents’ adverse drug reactions (ADRs).

Animals

Seven healthy adult Thoroughbreds.

Procedure

Mosapride 1.0 mg/kg and 2.0 mg/kg, metoclopramide 0.2 mg/kg, and cisapride 1.0 mg/kg were dissolved in 100 mL distilled water for oral administration. Lidocaine 1.3 mg/kg was mixed with 500 mL saline for a 30-min intravenous infusion. Oral administration of 100 mL distilled water was used as control. Gastric emptying was evaluated using ¹³CO₂ breath test, and jejunal and caecal motility was assessed by electrointestinography.

Results

The present study demonstrates that mosapride at doses of 1.0 mg/kg and 2.0 mg/kg facilitates gastric emptying in horses. Improved jejunal motility was observed following administration of mosapride (1.0 mg/kg and 2.0 mg/kg), metoclopramide (0.2 mg/kg), and cisapride (1.0 mg/kg). Similarly, improved caecal motility was observed following administration of mosapride (2.0 mg/kg).

Conclusions and clinical relevance

This study shows that among the prokinetic agents studied here, only mosapride (2.0 mg/kg) promotes jejunal and caecal motility in horses. Considering mosapride ADRs profile, it is believed that this compound is useful in the treatment of diseases associated with decreased GI motility, including postoperative ileus.     

The Effectiveness of Immunotherapy in Treating Exercise-Induced Pulmonary Hemorrhage

Inflammatory airway disease has been linked to exercise-induced pulmonary hemorrhage (EIPH), and consequently, we hypothesized that immunomodulation via concentrated equine serum (CES) treatment would reduce EIPH as evidenced by red blood cell (RBC) concentrations in bronchoalveolar lavage fluid (BALF). Separate trials were conducted on Thoroughbred horses treated with either CES (n = 6) or placebo (0.9% saline; n = 4). All horses completed pre-treatment and post-treatment (2 and 4 weeks after initiating treatment) maximal exercise tests on a 10% inclined treadmill (1 m/s/min increments to fatigue) over a 10-week period (2−3 weeks between tests), with bronchoalveolar lavage (BAL) performed 30 minutes after exercise. Treatment ensued 10 days after the pre-treatment exercise test, with horses receiving a series of five CES or placebo injections 24 hours apart (20 mL intratracheal and 10 mL intravenously), with subsequent weekly injections for 5 weeks thereafter. After CES treatment, both EIPH (RBC in BALF) and inflammation (white blood cell concentration [WBC] in BALF) were significantly diminished by the 4-week posttreatment run, demonstrating 46 ± 12% and 24 ± 11% decreases, respectively (P < 0.05). In contrast, EIPH was elevated significantly at the 4-week time point, and inflammation remained constant in the placebo trial. In conclusion, these preliminary data suggest that therapeutic intervention involving immunomodulation may represent a viable approach to reducing the severity of EIPH.     

Ghrelin differentially modulates the GH secretory response to GHRH between the fed and fasted states in sheep

The effect of energy balance on the growth hormone (GH) secretory responsiveness to growth hormone-releasing hormone (GHRH) has not been determined in ruminant animals. Therefore, we examined the effects of intravenous injections of 0, 3.3, and 6.6 μg ghrelin/kg body weight (BW), with and without GHRH at 0.25 μg/kg BW, on GH secretory responsiveness in both the fed and fasted sheep. The injections were carried out at 48 h (Fasting state) and 3 h (Satiety state) after feeding. Blood samples were taken every 10 minutes, from 30 minutes before to 120 minutes after the injection. Low (3.3 μg/kg BW) and high (6.6 μg/kg BW) doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Growth hormone-releasing hormone plus both doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Ghrelin and GHRH exerted a synergistic effect in the Satiety state, but not in the Fasting state. Plasma ghrelin levels were maintained significantly (P < .05) greater in the Fasting state than in the Satiety state except the temporal increases after ghrelin administration. Plasma free fatty acid (FFA) concentrations were significantly (P < .01) greater in the Fasting state than in the Satiety state. In conclusion, the present study has demonstrated for the first time that ghrelin differentially modulates GH secretory response to GHRH according to feeding states in ruminant animals.