Pharmacokinetics/pharmacodynamics cut-off determination for fosfomycin using Monte Carlo simulation in healthy horses.

Fosfomycin (FOM) is an approved veterinary medicinal product for large animals in Japan, but Clinical breakpoint (CBP) for antimicrobial susceptibility test (AST) is not defined for animals. This study aimed at conducting a pharmacokinetics/pharmacodynamics (PK/PD) analysis to determine the PK/PD cutoff for the CBP in horses. Drug concentrations following single intravenous administration (IV) of 20 mg/kg body weight (BW) FOM in nine horses were measured using liquid chromatography/mass spectrometry. The data were modelled using a nonlinear mixed-effects model, followed by Monte Carlo simulations. A 90% probability of target attainment for a PK/PD target of the ratio of Area Under the free plasma concentration-time curve divided by the minimal inhibitory concentration (MIC) >24 hr was set as PK/PD cut-off. The PK/PD cutoff for FOM 20 mg/kg BW q12 hr IV was estimated with the MIC value of ≤16.0 mg/L, and this regimen was considered effective against E. coli (MIC90; 16.0 mg/L) in healthy horses based on the MIC90 values of the wild population. Owing to the relevance of FOM to human health, veterinarians should use q 12 hr FOM 20 mg /kg against E. coli infections with an MIC <16 µg/mL, as suggested by our PK/PD cutoff after AST.

Evaluation of plasma and urine pharmacokinetics of tranexamic acid for equine medication control.

This study aimed to evaluate the pharmacokinetics (PK) of tranexamic acid (TXA) in horses and estimate its irrelevant plasma and urine concentrations using the pharmacokinetic/pharmacodynamic (PK/PD) approach by applying the Pierre-Louis Toutain model. TXA was intravenously administered to eight thoroughbred mares, and plasma and urine TXA concentrations were quantified by liquid chromatography/tandem mass spectrometry. The quantified data were used to calculate the PK parameters of TXA in horses. The plasma elimination curves were best-fitted to a three-compartment model. Using the Toutain model approach, irrelevant plasma and urine TXA concentrations were estimated to be 0.0206 and 0.997 µg/mL, respectively. The typical values of clearance, steady-state volume of distribution, and steady-state urine-to-plasma ratio were 0.080 L/kg/h, 0.86 L/kg, and 49.0, respectively. The obtained irrelevant concentrations will be useful for establishing relevant regulatory screening limits for effective control of TXA use in horse racing and equestrian sports.

Bacterial and fungal isolates from 107 cases of ulcerative keratitis in Japanese Thoroughbred racehorses (2017-2021).

Infectious ulcerative keratitis is a common disease in racehorses. To improve treatment outcomes, this study aimed to assess the antimicrobial susceptibilities of bacterial and fungal isolates obtained from the cornea of Japanese Thoroughbred racehorses with equine infectious ulcerative keratitis. Bacterial and fungal cultures were performed for 166 corneal swabs from 107 cases. A disc diffusion test and minimum inhibitory concentration test were also performed to assess antimicrobial susceptibility of the bacterial and fungal isolates, respectively. Bacterial and/or fungal isolates were obtained from 85.0% (91/107) of the cases. Staphylococcus was primarily isolated from bacterial isolates, including methicillin-resistant Staphylococcus aureus (MRSA), Aerococcus, Streptococcus, Acinetobacter, and Pseudomonas. Aspergillus was primarily isolated from filamentous fungi, and Debaryomyces species was primarily identified in yeast-like fungi. Ofloxacin resistance was observed in 100% (12/12), 15.9% (7/44), and 25.0% (3/12) of MRSA, Staphylococcus, and Streptococcus isolates, respectively. The prevalence of quinolone-resistant Staphylococci and Streptococci has increased in the past two decades. All Aspergillus isolates were susceptible to voriconazole, whereas other filamentous fungi, including Fusarium, were less susceptible to voriconazole. Further studies are required to determine effective treatments for antimicrobial-resistant isolates.

Pharmacokinetic-pharmacodynamic cutoff values for benzylpenicillin in horses to support the establishment of clinical breakpoints for benzylpenicillin antimicrobial susceptibility testing in horses.

Introduction: The aim of this international project was to establish a species-specific Clinical Breakpoint for interpretation of Antimicrobial Susceptibility Testing of benzylpenicillin (BP) in horses. Methods: A population pharmacokinetic model of BP disposition was developed to compute PK/PD cutoff values of BP for different formulations that are commonly used in equine medicine around the world (France, Sweden, USA and Japan). Investigated substances were potassium BP, sodium BP, procaine BP, a combination of procaine BP and benzathine BP and penethamate, a prodrug of BP. Data were collected from 40 horses that provided 63 rich profiles of BP corresponding to a total of 1022 individual BP plasma concentrations. Results: A 3-compartment disposition model was selected. For each of these formulations, the PK/PD cutoff was estimated for different dosage regimens using Monte Carlo simulations. The fAUC/MIC or fT>MIC were calculated with a free BP fraction set at 0.4. For fAUC/MIC, a target value of 72 h (for a 72h treatment) was considered. For fT>MIC, efficacy was assumed when free plasma concentrations were above the explored MIC (0.0625-2 mg/L) for 30 or 40 % of the dosing interval. For continuous infusion, a fT>MIC of 90 % was considered. It was shown that a PK/PD cutoff of 0.25 mg/L can be achieved in 90 % of horses with routine regimen (typically 22,000 IU/kg or 12.4 mg/kg per day) with IM procaine BP once a day (France, Japan, Sweden but not USA1) and with IM sodium BP at 14.07 mg/kg, twice a day or IV sodium BP infusion of 12.4 mg/kg per day. In contrast, penethamate and the combination of procaine BP and benzathine BP were unable to achieve this PK/PD cutoff not even an MIC of 0.125 mg/L. Discussion: The PK/PD cutoff of 0.25 mg/L is one dilution lower than the clinical breakpoint released by the CLSI (0.5 mg/ L). From our simulations, the CLSI clinical breakpoint can be achieved with IM procaine BP twice a day at 22,000 IU i.e. 12.4 mg/kg.

Incidence of surgical site infection after internal fixation of the first phalangeal bone and the third metacarpal/metatarsal bone fractures in Thoroughbred racehorses.

Surgical site infection (SSI) is one of the major complications of equine fracture surgery. The purpose of this study was to investigate the incidence of and risk factors for SSI after internal fixation of the first phalangeal bone (P1) and the third metacarpal/metatarsal bone (MC3/MT3) fractures in Thoroughbred racehorses. Between 2011 and 2020, 451 cases underwent surgery with screws or a locking compression plate (LCP) for sagittal fractures of P1 or condylar fractures of MC3/MT3. Overall, 2.9% (13/451) of the cases developed an SSI. The incidence was significantly higher in plate fixation (21.4%) than in screw fixation (2.3%). There was no significant association with other variables, such as sex, age, number of screws, experience of surgeon, or prophylactic antimicrobials. The median duration of hospitalization for screw fixation was 14 days without an SSI and 20 days with an SSI, and those for plate fixation were 26 and 25-88 days, respectively, indicating that the development of SSI prolongs the duration of hospitalization. On the other hand, there were no significant differences in discharge and race resumption rates between cases with and without an SSI. These data indicate that the incidence of SSI in this study was low and that it was higher following plate fixation than screw fixation.

Characteristic Inflammatory Biomarkers in an Equine Model of Persistent Synovitis Induced By the Intra-Articular Administration of Monoiodoacetic Acid.

Persistent synovitis damages the articular cartilage in horses. To evaluate the effectiveness of treatment for synovitis using a model induced by intra-articular administration of monoiodoacetic acid (MIA), it is necessary to identify inflammatory biomarkers characteristic of the MIA model. Synovitis was induced by administering MIA into the unilateral antebrachiocarpal joints of five horses, and saline was injected into the contralateral joints as a control on day 0. Clinical and ultrasonographic examinations and synovial fluid collection were performed on days 0, 1, 2, 7, 14, 21, 28, and 35. Leukocyte, lactate dehydrogenase (LDH), tumor necrosis factor-α (TNF-α), interleukin-1 receptor antagonist (IL-1Ra), interleukin-6 (IL-6), and transforming growth factor-ß1 (TGF-ß1) concentrations in the synovial fluid were measured. Synovium was obtained after euthanasia on day 42 and histologically examined before quantification of the gene expression of inflammatory biomarkers by real-time PCR. Acute inflammatory symptoms persisted for approximately 2 weeks before returning to control levels. However, some indicators of chronic inflammation remained elevated until day 35. On day 42, synovitis continued histologically, with osteoclasts. The expressions of matrix metalloproteinase 13 (MMP13), a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4), receptor activator of nuclear factor kappa-Β ligand (RANKL), and collagen type I α2 chain (Col1a2) were significantly higher in the MIA model than in the control. In the MIA model, representative inflammatory biomarkers in the chronic inflammatory stage were persistently expressed in both synovial fluid and tissue, suggesting that they may be useful for the assessment of the anti-inflammatory effect of drugs.

Rational determination of cefazolin dosage regimen in horses based on pharmacokinetics/pharmacodynamics principles and Monte Carlo simulations.

A pharmacokinetics/pharmacodynamics (PK/PD) approach was used to determine the best empirical dosage regimen of cefazolin (CEZ) after intramuscular (IM) administration of CEZ in horses. Seven horses received a single IM or intravenous (IV) administration of CEZ of 5 mg/kg bodyweight (BW) according to a crossover design. CEZ plasma concentrations were measured using LC-MS/MS. The plasma concentrations in these seven horses and those of six other horses obtained in a previous study with an IV CEZ dose of 10 mg/kg were modelled simultaneously using NonLinear Mixed-Effect modelling followed by Monte Carlo simulations to establish a rational dosage regimen. A 90% Probability of Target Attainment (PTA) for a PK/PD target of a free plasma concentration exceeding MIC90 (fT > MIC ) for 40% of the dosing interval was set for selecting an effective dosing regimen. The typical half-life of absorption and bioavailability after IM administration were 1.25 h and 96.8%, respectively. A CEZ dosage regimen of 5 mg/kg BW q12h IM administration achieved therapeutic concentrations to control both Streptococcus zooepidemicus and Staphylococcus aureus. For the same dose, the fT > MIC after IM administration was significantly longer than after IV administration, and the IM route should be favoured by clinicians for its efficiency and convenience.

Pharmacokinetic/pharmacodynamic analysis of cephalothin after intramuscular administration in Thoroughbred horses.

A pharmacokinetic/pharmacodynamic (PK/PD) approach was used to determine a dosage regimen of cephalothin (CET) after intramuscular (IM) administration in horses. CET plasma concentrations were measured in eight horses after a single IM administration of 11 mg/kg bwt of CET. The data were modeled using a nonlinear mixed-effect model, and the probability of target attainment (PTA) of the PK/PD target was calculated for 5,000 horses generated by Monte Carlo simulations. IM administrations of CET at 11 mg/kg bwt q 8 hr and q 6 hr achieved a PTA of 90% against the MIC90 of S. zooepidemicus and S. aureus, respectively, and were considered to be effective dosage regimens. The total dose for the IM administration recommended in this study was lower than that for intravenous (IV) administration in previous studies.

Concentration of cephalothin in body fluids and tissue samples of Thoroughbred horses.

Cephalothin (CET) concentrations in body fluids (plasma, synovial fluid, pleural fluid, peritoneal fluid, and aqueous humor) and tissue samples (bone, lung, jejunum, hoof, and subcutaneous tissue) were investigated to consider the treatment of infectious diseases in horses. CET 22 mg/kg body weight was intravenously administered to 12 horses. Samples were collected from four different horses at 1, 3, and 5 hr after administration. The CET concentration in body fluids other than aqueous humor was maintained above the MIC90 values of Streptococcus zooepidemicus and Staphylococcus aureus until 5 hr, but it was not maintained above that of S. aureus in bone. CET (22 mg/kg twice a day) is effective for septic arthritis, pleuritis, and peritonitis caused by gram-positive bacteria but ineffective for osteomyelitis.

Generic approach for the discovery of drug metabolites in horses based on data-dependent acquisition by liquid chromatography high-resolution mass spectrometry and its applications to pharmacokinetic study of daprodustat.

In drug metabolism studies in horses, non-targeted analysis by means of liquid chromatography coupled with high-resolution mass spectrometry with data-dependent acquisition (DDA) has recently become increasingly popular for rapid identification of potential biomarkers in post-administration biological samples. However, the most commonly encountered problem is the presence of highly abundant interfering components that co-elute with the target substances, especially if the concentrations of these substances are relatively low. In this study, we evaluated the possibility of expanding DDA coverage for the identification of drug metabolites by applying intelligently generated exclusion lists (ELs) consisting of a set of chemical backgrounds and endogenous substances. Daprodustat was used as a model compound because of its relatively lower administration dose (100 mg) compared to other hypoxia-inducible factor stabilizers and the high demand in the detection sensitivity of its metabolites at the anticipated lower concentrations. It was found that the entire DDA process could efficiently identify both major and minor metabolites (flagged beyond the pre-set DDA threshold) in a single run after applying the ELs to exclude 67.7-99.0% of the interfering peaks, resulting in a much higher chance of triggering DDA to cover the analytes of interest. This approach successfully identified 21 metabolites of daprodustat and then established the metabolic pathway. It was concluded that the use of this generic intelligent "DDA + EL" approach for non-targeted analysis is a powerful tool for the discovery of unknown metabolites, even in complex plasma and urine matrices in the context of doping control.

Protective efficacy of a reverse genetics-derived inactivated vaccine against equine influenza virus in horses.

Updating vaccine strains is essential to control equine influenza. We evaluated the protective efficacy of an inactivated equine influenza vaccine derived from viruses generated by reverse genetics (RG) in horses in an experimental viral challenge study. Wild-type (WT) virus (A/equine/Tipperary/1/2019) and virus generated by RG (consisting of hemagglutinin and neuraminidase genes from A/equine/Tipperary/1/2019 and six other genes from high-growth A/Puerto Rico/8/34) were inactivated by formalin for vaccine use. Twelve 1-year-old naïve horses with no antibodies against equine influenza virus were assigned to three groups (each n = 4): control, WT, and RG. They were vaccinated twice, 4 weeks apart, and were challenged with A/equine/Tipperary/1/2019 2 weeks after the second vaccination. All four horses in the control group and one horse in the WT group had pyrexia for multiple days and respiratory illness, and one horse in the RG group had pyrexia for 2 days without respiratory illness. The mean rectal temperatures and the mean concentrations of serum amyloid A in the WT and RG groups were significantly lower than those in the control group, with no significant differences between them. The WT and RG vaccines significantly reduced viral shedding relative to the control. The protective efficacy of the RG-derived inactivated vaccine against equine influenza virus is comparable to that of the vaccine derived from WT viruses in horses. The RG technique can make it easy to update equine influenza vaccine strains.

Pharmacokinetic Study of Vadadustat and High-Resolution Mass Spectrometric Characterization of its Novel Metabolites in Equines for the Purpose of Doping Control.

BACKGROUND: Vadadustat, a hypoxia-inducible factor prolyl hydroxylase (HIF-PHD) inhibitor, is a substance which carries a lifetime ban in both horse racing and equestrian competition. A comprehensive metabolic study of vadadustat in horses has not been previously reported. OBJECTIVE: Metabolism and elimination profiles of vadadustat in equine plasma and urine were studied for the purpose of doping control. METHODS: A nasoesophageal administration of vadadustat (3 g/day for 3 days) was conducted on three thoroughbred mares. Potential metabolites were comprehensively detected by differential analysis of full-scan mass spectral data obtained from both in vitro studies with liver homogenates and post-administration samples using liquid chromatography high-resolution mass spectrometry. The identities of metabolites were further substantiated by product ion scans. Quantification methods were developed and validated for the establishment of the excretion profiles of the total vadadustat (free and conjugates) in plasma and urine. RESULTS: A total of 23 in vivo and 14 in vitro metabolites (12 in common) were identified after comprehensive analysis. We found that vadadustat was mainly excreted into urine as the parent drug together with some minor conjugated metabolites. The elimination profiles of total vadadustat in post-administration plasma and urine were successfully established by using quantification methods equipped with alkaline hydrolysis for cleavage of conjugates such as methylated vadadustat, vadadustat glucuronide, and vadadustat glucoside. CONCLUSION: Based on our study, for effective control of the misuse or abuse of vadadustat in horses, total vadadustat could successfully be detected for up to two weeks after administration in plasma and urine.

Outbreak of methicillin-resistant Staphylococcus aureus sequence type 1, spa type t1784, in an equine hospital in Japan.

Methicillin-resistant Staphylococcus aureus (MRSA) has often been isolated from livestock and companion animals, including horses. Seven cases of MRSA infection in Thoroughbred racehorses were observed in an equine hospital in Japan in 2020. In this study, MRSA isolates from these seven horses and nine veterinarians in the equine hospital were studied to examine their genetic relatedness and evaluate the possibility of MRSA transmission. The MRSA isolates were subjected to whole-genome sequencing for multi-locus sequence typing, S. aureus protein A (spa) typing, staphylococcal cassette chromosome typing, and antimicrobial resistance gene detection. Minimum inhibitory concentrations of antibiotics were assessed to determine the antimicrobial susceptibility phenotype of the isolates. Phylogenetic trees based on single nucleotide polymorphisms were constructed to identify genetically close isolates. All isolates from horses and veterinarians belonged to sequence type (ST) 1, spa type t1784, with a point mutation in gyrA and double point mutations in grlA, which is known to cause fluoroquinolone resistance. All ST1-t1784 isolates were genetically closely related based on the phylogenetic tree. Our results suggested an outbreak and horse-veterinarian transmission of ST1-t1784 strains in an equine hospital.

Incidence of carpal fractures and risk factors for recurrent fractures after arthroscopic removal of carpal chip fracture fragments in Thoroughbred racehorses.

BACKGROUND: We aimed to investigate the recent incidence of carpal fractures and the risk factors for recurrent ipsilateral fractures after arthroscopic removal of clinically active unilateral carpal chip fracture fragments in Thoroughbred racehorses. METHODS: The findings for horses managed under the Japan Racing Association that developed carpal bone fractures between 2014 and 2018 were retrospectively reviewed. The proportion of cases that developed a recurrent carpal fracture in the originally affected joint was calculated, and the risk factors for recurrent fractures were analysed. RESULTS: In total, 2858 carpal fractures were recorded in the study period (incidence, 0.8%). Of the 554 horses that resumed racing after the treatment of the unilateral major carpal chip fracture, 144 had a recurrent fracture (26.0%). Chip fractures of the third carpal bone (odds ratio [OR]: 3.7) or a combination of the distal end of the radius and intermediate carpal bone (OR: 3.0) were associated with a significantly higher risk of recurrent fractures than the distal aspect of the radial carpal bone. CONCLUSIONS: The incidence of carpal fractures remained similar to that reported in Japan in the 1990s. The rate of recurrent ipsilateral fractures differed among lesion sites.

Platelet Lysate Enhances Equine Skeletal Muscle Regeneration in A Bupivacaine-Induced Muscle Injury Model☆.

This study aimed to verify the effects of platelet lysate (PL) administration on the repair of injured horse tissue. Skeletal muscle injuries were induced in 26 Thoroughbreds by bupivacaine administration. PL or saline was administered 1 day (1D) after injury. Muscle samples from 22 horses injected with PL or saline were obtained by needle biopsy at 2, 3, 4, or 7 days (2D, 3D, 4D, or 7D, respectively) after injury, and growth-factor concentrations and muscle regeneration-associated gene expression levels were determined. Intact samples were similarly collected before injury, and samples of injured muscle not treated with PL or saline (sham samples) were also obtained at 1D, 2D, 3D, 4D, and 7D as references for comparison. Samples from the remaining 4 horses were obtained by surgical incision following euthanasia at 5 days (5D) and 7D after injury, followed by histological analysis. Although increased growth factor levels caused by PL administration were observed for up to 1-day post-administration (2D), gene expressions were enhanced for up to 6 days post-administration (7D). Moreover, the number of embryonic myosin heavy chain (MHC-e)-positive myofibrils at 5D was higher in the PL-treated group than in the saline-treated group, whereas no significant between-group difference in the number of myofibrils was recorded at 7D. Thus, PL administration in muscle injury upregulated the expression of various genes associated with muscle regeneration and promoted morphological regeneration within 6 days of treatment, although growth factor levels from PL decreased at the injected site by approximately 2 days post-administration.

A comprehensive and comparative proteomic analysis of horse serum proteins in colitis.

BACKGROUND: Equine colitis is a diarrhoeal disease caused by inflammation of the large bowel and can potentially be life-threatening due to its rapid progression. Pathogenesis is multifactorial and pathophysiology is highly complicated, therefore, reliable diagnostic biomarkers are needed in the veterinary field. OBJECTIVE: Serum is one of the most commonly used diagnostic tools in equine clinical investigation. To discover diagnostic or prognostic protein markers for colitis in horse serum, comprehensive and comparative proteomic analysis was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS). STUDY DESIGN: Case-control study. METHODS: Serum samples were collected from 36 healthy Thoroughbreds and 12 Thoroughbreds with colitis. Serum from each horse suffering from colitis was collected daily until death or recovery. Collected sera were digested with trypsin. Peptides obtained from serum proteins were measured by Q-Exactive HF Orbitrap mass spectrometer. The identification and quantification of peptides were performed using Proteome Discoverer version 2.2. RESULTS: On day 1 of treatment, eight proteins in the colitis group were upregulated (P < .05, more than a twofold change) compared with the healthy group. Among the eight proteins, biliverdin reductase B was significantly upregulated (P < .05) in the non-survivor group (n = 5) compared with the survivor group (n = 7). On the last day of the treatment, haemoglobin subunit alpha, clusterin, glyceraldehyde-3-phosphate dehydrogenase, lipopolysaccharide-binding protein, and biliverdin reductase B showed significant increases (P < .05) in the non-survivor group. MAIN LIMITATIONS: The number of the identified proteins is limited due to the existence of abundant proteins. CONCLUSIONS: Measuring the changes of these proteins together may enable a potential prognosis or early diagnosis of horses suffering from colitis.

Medication control of flunixin in racing horses: Possible detection times using Monte Carlo simulations.

BACKGROUND: For medication control in several jurisdictions, withdrawal time is the period of refrain from racing after drug administration. It is set by adding a safety period to an experimental detection time. However, there are no reports of statistical analyses of detection time for the determination of withdrawal time in flunixin meglumine-treated horses. OBJECTIVE: To analyse the population pharmacokinetics of flunixin in horses through the generation of a dataset for detection time statistical analysis and predictions via Monte Carlo simulation. STUDY DESIGN: Experimental study. METHODS: Drug plasma and urine concentrations following single intravenous administration of flunixin 1.1 mg/kg bodyweight (BW) in 10 horses and multiple administration of q 24 hours for 5 days in 10 horses were measured using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Data were modelled using a nonlinear mixed effect model followed by Monte Carlo simulation. Irrelevant plasma concentration (IPC) and irrelevant urine concentration (IUC) were calculated using the Toutain approach. Detection times were obtained considering the time after the last administration for selected quantiles of 5000 hypothetical horses under the international screening limit (ISL) proposed by the International Federation of Horseracing Authorities (plasma: 1 ng/mL, urine; 100 ng/mL). RESULTS: For a regimen of 1.1 mg/kg BW q 24 hours, the IPC and IUC values were 2.0 and 73.0 ng/mL respectively. Detection times in plasma above the ISL for 90% of simulated horses were estimated as 74 hours after a single 1.1 mg/kg dose administration, 149 and 199 hours after multiple doses over 5 days at either 24- or 12-hour intervals respectively. Corresponding detection times in urine were 46, 68 and 104 hours respectively. MAIN LIMITATION: Only female horses were investigated. CONCLUSIONS: Statistical detection times for different flunixin meglumine regimens indicated a delay of detection time in plasma after multiple administrations under ISL.

Rational dosage regimens for cephalothin and cefazolin using pharmacokinetics and pharmacodynamics analysis in healthy horses.

BACKGROUND: First-generation cephalosporins have good activity against gram-positive bacteria and are extensively used in horses. There are few reports of pharmacokinetics and pharmacodynamics (PK/PD) analysis of cephalosporins in horses. OBJECTIVE: To optimise the dosages of the two first-generation cephalosporins cephalothin (CET) and cefazolin (CEZ) in horses using PK/PD concepts. STUDY DESIGN: Experimental study with single administration. METHODS: Drug plasma concentrations following a single intravenous (i.v.) administration of 22 mg/kg bodyweight (bwt) CET in 12 horses and of 10 mg/kg bwt CEZ in six horses were measured using LC-MS/MS. Data were modelled using a nonlinear mixed effect modelling followed by Monte Carlo simulations. Minimum inhibitory concentrations (MICs) against Streptococcus zooepidemicus and Staphylococcus aureus isolated from horses were determined by the microbroth dilution method. RESULTS: The percentages of CET and CEZ binding to serum proteins were 19.9% ± 8.4% and 15.2% ± 8.5% respectively. For both CET and CEZ, the MIC90 against S. zooepidemicus was 0.12 mg/L and against S. aureus was 0.5 mg/L. For CET, to achieve a probability of target attainment (PTA) of 90% for a PK/PD target of a free serum plasma concentration exceeding the MIC90 for 40% of the dosing interval, an empirical CET dosage regimen of 22 mg/kg bwt q8h and 22 mg/kg bwt q4h i.v. administration were required for S. zooepidemicus and S. aureus respectively. For CEZ, the corresponding dosage regimens were 10 mg/kg bwt q12h and 10 mg/kg bwt q8h. MAIN LIMITATIONS: Small sample size only in healthy horses. CONCLUSIONS: For CET, more frequent administration than that currently recommended (22 mg/kg bwt q6-12h) is required to empirically control S. aureus infection in horses. For CEZ, less frequent administration compared to the dosage regimen currently proposed (10-22 mg/kg bwt q6h) could control S. zooepidemicus and S. aureus infections in horses.

Optimal double-spin method for maximizing the concentration of platelets in equine platelet-rich plasma.

This study optimized the double-spin conditions for preparing equine platelet-rich plasma (PRP): leukocyte-rich PRP (L-PRP) and leukocyte-poor PRP (P-PRP). Whole blood samples were centrifuged at various double-spin conditions. Both L-PRP and P-PRP were prepared at each stage, and complete blood counts and growth factor concentrations were compared. Samples centrifuged at 160 × 900 g, 160 × 2,000 g, and 400 × 2,000 g exhibited the highest platelet counts. P-PRP had significantly lower leukocyte and erythrocyte contents than L-PRP, especially at 400 × 2,000 g. No significant differences were observed in growth factor concentrations. Our data suggest that optimum L-PRP preparation should include centrifugation under the aforementioned conditions, whereas centrifugation at 400 × 2,000 g is optimal for P-PRP.

Single-dose pharmacokinetics of orally administered metronidazole and intravenously administered imipenem in healthy horses and computer-based simulation of pleural fluid concentrations with multiple dosing.

OBJECTIVE: To determine plasma pharmacokinetics of metronidazole and imipenem following administration of a single dose PO (metronidazole, 15 mg/kg) or IV (imipenem, 10 mg/kg) in healthy Thoroughbreds and simulate pleural fluid concentrations following multiple dose administration every 8 hours. ANIMALS: 4 healthy Thoroughbreds. PROCEDURES: Metronidazole and imipenem were administered, and samples of plasma and pleural fluid were collected at predetermined time points. Minimum concentrations of metronidazole and imipenem that inhibited growth of 90% of isolates (MIC90), including 22 clinical Bacteroides isolates from horses with pleuropneumonia, were calculated. For the computer simulation, the target ratio for area under the pleural fluid concentration-versus-time curve during 24 hours to the MIC90 for metronidazole was > 70, and the target percentage of time per day that the pleural fluid concentration of imipenem exceeded the MIC90 was > 50%. RESULTS: Mean ± SD pleural fluid concentrations of metronidazole and imipenem were 12.7 ± 3.3 µg/mL and 12.1 ± 0.9 µg/mL, respectively, 1 hour after administration and 4.9 ± 0.85 µg/mL and 0.3 ± 0.08 µg/mL, respectively, 8 hours after administration. For both antimicrobials, concentrations in the pleural fluid and plasma were similar. The ratio for area under the pleural fluid concentration-versus-time curve during 24 hours to the MIC90 for metronidazole was 84.9, and the percentage of time per day the pleural fluid concentration of imipenem exceeded the MIC90 was 70.9%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that administration of metronidazole (15 mg/kg, PO, q 8 h) or imipenem (10 mg/kg, IV, q 8 h) resulted in their accumulation in the pleural fluid in healthy horses and concentrations were likely to be effective for the treatment of pneumonia and pleuropneumonia caused by Bacteroides spp.