Determination of intracellular concentrations of free and two types of liposome-encapsulated enrofloxacin in anatolian shepherd dog monocytes

In this study, it was evaluated the accumulation of free and two types of liposome-encapsulated enrofloxacin (LEE) at the doses of 0.25, 0.5 and 1 microg/ml, which were clinically relevant concentrations into monocytes of healthy Anatolian shepherd dogs. Enrofloxacin was encapsulated with two different types of liposome in multilamellar large vesicles (MLV). Type A MLV composed of 15 mg egg phosphatidylcholine and 35 mg cholesterol, Type B MLV composed of phosphatidylcholine (PC), cholesterol and enrofloxacin, in a molar ratio of 1 : 1 : 1. The mean sizes of Type A and Type B liposome were found to be 7.65 and 4.27 microm, respectively. However, the mean encapsulation rate determined of Type A (13 +/- 2%) was found lower than Type B liposome (44 +/- 3%). The amounts of intracellular enrofloxacin concentrations were determined by high performance liquid chromatography. Type B LEE accumulated significantly higher level into monocytes when compared to free drug or Type A liposome. This study showed that Type B LEE markedly concentrated within monocytes and may improve the antibacterial efficacy of the antibiotic.     

Determination of Intracellular Concentrations of Free and Two Types of Liposome‐Encapsulated Enrofloxacin in Anatolian Shepherd Dog Monocytes

In this study, it was evaluated the accumulation of free and two types of liposome‐encapsulated enrofloxacin (LEE) at the doses of 0.25, 0.5 and 1 μg/ml, which were clinically relevant concentrations into monocytes of healthy Anatolian shepherd dogs. Enrofloxacin was encapsulated with two different types of liposome in multilamellar large vesicles (MLV). Type A MLV composed of 15 mg egg phosphatidylcholine and 35 mg cholesterol, Type B MLV composed of phosphatidylcholine (PC), cholesterol and enrofloxacin, in a molar ratio of 1 : 1 : 1. The mean sizes of Type A and Type B liposome were found to be 7.65 and 4.27 μm, respectively. However, the mean encapsulation rate determined of Type A (13 ± 2%) was found lower than Type B liposome (44 ± 3%). The amounts of intracellular enrofloxacin concentrations were determined by high performance liquid chromatography. Type B LEE accumulated significantly higher level into monocytes when compared to free drug or Type A liposome. This study showed that Type B LEE markedly concentrated within monocytes and may improve the antibacterial efficacy of the antibiotic.     

Fragment Bb of bovine complement factor B: stimulatory effect on the microbicidal activity of bovine monocytes.

We have previously shown that the Bb fragment of bovine complement factor B activates bovine monocytes and neutrophils. The activation was demonstrated by the enhanced uptake of 3H-deoxyglucose. To investigate the potential effect of fragment Bb on the microbicidal activity of bovine monocytes, a direct method was used. This method involves an initial ingestion period at 37 degrees C followed by repeated washing. The decrease in the total number of viable intracellular Staphylococcus aureus during the reincubation of the bacteria with bovine monocytes determines the intracellular killing. Maximal intracellular killing was seen when the monocytes containing the ingested S. aureus was incubated with fresh bovine serum (mean +/- SEM = 73.4 +/- 1.4%). On incubation of the monocytes, containing the ingested bacteria with heat-inactivated bovine serum, 32.5 +/- 0.7% of the intracellular bacteria were killed. When affinity-purified bovine factor Bb was added to the heat-inactivated serum, the intracellular killing capacity was almost restored (65.8 +/- 1.5%). When monocytes were incubated with medium alone, they killed 22.4% of the intracellular microorganisms. When fragment Bb (25 micrograms/mL) was added to the medium, the intracellular killing of S. aureus doubled (46 +/- 1.29%). We conclude that the Bb fragment of bovine complement factor B stimulates bovine monocytes in their microbicidal activity.     

The mechanism of enhanced intraphagocytic killing of bacteria by liposomes containing antibiotics

Liposomes containing aminoglycosides have been shown to enhance the killing of Brucella abortus and Staphylococcus aureus inside bovine phagocytic cells. This study examined the mechanism by which liposomes containing aminoglycoside enhance the intracellular killing of bacteria. Liposomes with entrapped aminoglycoside were found to significantly enhance the intraphagocytic killing of bacteria in bovine phagocytic cells (in vitro) when compared to free drug. Liposomes with entrapped aminoglycoside were also found to deliver significantly higher levels of aminoglycoside into phagocytic cells when compared to free drug (gentamicin) or free drug and liposomes without entrapped antibiotic. Antibiotic delivered to adherent phagocytic cells could be detected 3 days after treatment of the cells with liposomes containing aminoglycoside. No antibiotic could be detected in the supernatants of phagocytic cell cultures 3 days after treatment with liposomes containing antibiotic was only observed when the intraphagocytic bacteria were sensitive to the antibiotic entrapped in the liposomes. The rate of phagocytosis of S. aureus by cells treated with cationic liposomes (no entrapped antibiotic) did not differ from the rate of phagocytosis of control cells not treated with cationic liposomes. This study shows that the enhanced intraphagocytic killing of bacteria in bovine phagocytic cells occurs by direct delivery of entrapped antibiotic into the phagocytic cell by the liposome delivery vehicle and not by nonspecific enhancement of phagocytic cell function. Liposomes containing aminoglycoside appear to have no toxic effects on phagocytic cell function or viability in vitro.     

The use of liposomally-entrapped gentamicin in the treatment of bovine Staphylococcus aureus mastitis.

The effect of incorporation of gentamicin in liposomes on intracellular killing of Staphylococcus aureus was studied in vitro in cultured bovine mammary macrophages, and in experimental bovine mastitis. Liposomes were prepared by reverse-phase evaporation and ranged in size from 0.1 to 1.0 micron in diameter (mean 0.51 micron), with an encapsulation efficiency of gentamicin of 27.4%. Liposomes were taken up by in vitro cultured macrophages but intracellular killing of S. aureus over 12 h was not significantly enhanced when treatment with liposomally-entrapped gentamicin was compared to free gentamicin. Treatment of experimentally-induced S. aureus mastitis in five lactating Holstein cows (20 quarters) failed to show significant differences in bacterial counts when treatment with liposomally-entrapped gentamicin was compared to treatment with free gentamicin or blank liposomes plus free gentamicin. Gentamicin concentrations exceeded the in vitro determined minimum inhibitory concentration for 48 h when quarters were treated with 50 mg gentamicin on two occasions 24 h apart.     

Bovine leukocyte phagocytosis and bacteria killing monitored by intracellular acridine orange fluorescence and extracellular fluorescence quenching

The time course of phagocytosis and intracellular killing of serum-opsonized Escherichia coli K12 and Staphylococcus aureus SG511 by glass-adherent bovine peripheral blood polymorphonuclear leukocytes (PMNLs) and cultured monocytes (macrophages) was monitored by fluorescence microscopy of single cells using the acridine orange (AO)/crystal violet (CV) technique. After interaction of glass-adherent leukocytes (20, 40, 60 min, 37 degrees C) with opsonized bacteria, cells were stained with the fluorescent dye AO. Living bacteria stained green, dead bacteria stained orange. The addition of CV to AO-stained bacteria quenched the fluorescence of extracellular bacteria only. CV does not penetrate living bovine PMNLs which allows the discrimination of ingested (fluorescent) and extracellular (nonfluorescent) bacteria during attachment and phagocytosis of bacteria by adherent PMNLs. We investigated quantitatively phagocytosis and intracellular killing of serum-opsonized bacteria by bovine PMNLs from 22 bulls of 4 different Swiss dairy breeds. Within 60 min maximum uptake (approximately 12 bacteria/PMNL) and killing (approximately 80%) of serum-opsonized Escherichia coli K12 and Staphylococcus aureus SG511 were achieved. The AO/CV technique was also used to quantify the uptake and intracellular killing of serum-opsonized Escherichia coli K12 by cultured monocytes (macrophages). Within 60 min maximum uptake of bacteria (approximately 16/MO) was achieved; approximately 83% of bacteria were killed.     

Intracellular killing of mastitis pathogens by penethamate hydriodide following internalization into mammary epithelial cells

Penethamate hydriodide was highly effective in killing Streptococcus uberis, Streptococcus dysgalactiae subsp. dysgalactiae and Staphylococcus aureus that internalized mammary epithelial cells. At higher concentrations (32 microg/mL to 32 mg/mL), killing rates ranged from 85% to 100%. At lower concentrations (0.032 microg/mL to 3.2 microg/mL), killing rates ranged from 0 to 80%. Results of this proof-of-concept study demonstrated that: (1) penethamate hydriodide is capable of entering mammary epithelial cells and killing intracellular mastitis pathogens without affecting mammary epithelial cell viability, (2) the in vitro model used is capable of quantifying the fate of mastitis pathogens internalized into mammary epithelial cells, and (3) this in vitro model can be used to determine the effectiveness of antibiotics at killing bacteria within the cytoplasm of mammary epithelial cells.     

Interaction between Salmonella typhimurium and phagocytic cells in pigs. Phagocytosis, oxidative burst and killing in polymorphonuclear leukocytes and monocytes

Interactions between Salmonella typhimurium and peripheral blood leucocytes from healthy, Salmonella-free pigs were investigated in vitro. Both granulocytes and monocytes phagocytized FITC-labelled heat-killed Salmonella bacteria as shown by flow cytometry. Phagocytosis in whole blood and isolated leucocytes was measured as acquired fluorescence in the leukocytes and was both time and dose related. Living, serum-opsonized Salmonella bacteria induced a dose-dependent oxidative burst in PMNs and monocytes as measured by luminol-enhanced chemiluminescence (LC). When opsonized in normal serum the Salmonella bacteria, in the range of 2-5 x 10(7) cfu, induced a LC response in monocytes comparable to the level of responses induced by phorbol myristate acetate (PMA) and opsonized zymosan, and the Salmonella-induced response was only marginally reduced by superoxide dismutase (SOD). Intracellular killing of Salmonella by monocytes was assessed from plate colony counts of lysed monocytes and showed that Salmonella typhimurium was able to survive and proliferate in adherent monocytes in vitro despite a reduction in intracellular cfu during the first hour's incubation in cells from some pigs. Experiments with the exhaustion of oxidative burst in non-adherent monocytes were performed by prestimulation with PMA, heat-killed Salmonella or buffer. Prestimulation with PMA led to a strong reduction in oxidative burst induced by living opsonized Salmonella bacteria, whereas prestimulation with heat-killed bacteria gave rise to an enhanced response. In these experiments intracellular killing of the added living Salmonella gave variable results, in that monocytes from two out of three pigs showed no essential change in intracellular bactericidal activity, but with cells from one pig a less pronounced bactericidal activity was found after prestimulation with PMA.     

Bovine pulmonary alveolar macrophages: antemortem recovery and in vitro evaluation of bacterial phagocytosis and killing

A system was developed to recover pulmonary alveolar macrophages (PAM) from living cattle and to evaluate the function of these cells by measuring bacterial phagocytosis and killing. For the collection of PAM, single-tube and telescoped double-tube pulmonary lavage devices were compared. The total recovery, using these systems, was 70 +/- 10.7% of infused fluid, yielding approximately 87% PAM. The total number of cells per collection was approximately 5 times higher with the single-tube device (6.87 +/- 0.78 x 10(7) cell/ml) than with the telescoped double-tube device (1.3 +/- 0.1 X 10(7) cells/ml). Phagocytosis and intracellular killing of Staphylococcus epidermidis and Staphylococcus aureus by PAM in media suspension and by plastic-adherent PAM were evaluated. In addition, different bacteria-to-macrophage ratios were assessed, as well as the intracellular killing of S epidermidis at periodic intervals. Results showed that over a 3-hour period, similar numbers of both bacteria were phagocytized, but intracellular killing of S epidermidis was more efficient than intracellular killing of S aureus. It also was found (i) that suspended PAM and adherent PAM phagocytized similar numbers of bacteria; (ii) that when the bacteria-to-cell ratio was 10:1, the numbers of phagocytized bacteria and intracellular killing were higher than when the ratio was 1:10; and (iii) that killing of S epidermidis by adherent PAM was directly proportional to incubation time. The time that PAM are in culture affects the phagocytosis and killing of intracellular bacteria, as shown by increased phagocytosis and by intracellular killing of S epidermidis by PAM in suspension for 48 hours or plastic adherent for 60 hours after collection.     

Effect of temperature modulation and bvg mutation of Bordetella bronchiseptica on adhesion, intracellular survival and cytotoxicity for swine alveolar macrophages

Bordetella bronchiseptica causes respiratory disease in swine, yet there are no studies examining the interaction of B. bronchiseptica with swine alveolar macrophages. A swine isolate of B. bronchiseptica was able to adhere to, and survive intracellularly in, swine alveolar macrophages, but the relative ability of the bacteria to accomplish these functions was dependent on its phenotypic phase and culture conditions. More bacteria were observed extracellularly as well as intracellularly by immunofluorescent staining when B. bronchiseptica was cultured at 23 degrees C as compared to 37 degrees C. However, more bacteria cultured at 37 degrees C were found surviving intracellularly after the macrophages were cultured with polymyxin B to kill extracellular bacteria. Similar results were seen in experiments performed with an isogenic Bvg(-) phase-locked mutant of B. bronchiseptica cultured at 37 or 23 degrees C, indicating that another temperature dependent mechanism in addition to bvg may play a role in adhesion and intracellular survival. B. bronchiseptica was cytotoxic for swine alveolar macrophages in the Bvg(+) phase only. The cytotoxicity of B. bronchiseptica for alveolar macrophages, and its ability to survive phagocytosis, are no doubt important to escape from immune clearance mechanisms and establish infection, and could leave the host susceptible to secondary respiratory pathogens.     

Antibacterial activity of marbofloxacin. A new fluoroquinolone for veterinary use against canine and feline isolates

Marbofloxacin is a new fluoroquinolone developed exclusively for veterinary use. Minimum inhibitory concentrations of marbofloxacin were assessed for 816 recent isolates associated with canine or feline diseases. Marbofloxacin showed a broad spectrum of activity against gram-negative and gram-positive bacteria. In vitro rates of killing of marbofloxacin and enrofloxacin were compared against strains of Staphylococcus intermedius and Pasteurella multocida , and the results showed no marked difference between the two antibiotics. The duration of bactericidal activity was evaluated ex vivo in the urine of dogs and cats treated with marbofloxacin and lasted from 2 to 5 days after a single administration according to the dosages. Post-antibiotic effect durations were determined with Escherichia colt, PasteureUa multocida, Staphylococcus aureus and Staphylococcus intermedius and were found almost equal to those of enrofloxacin or ciprofloxacin. These results predict a great potential for marbofloxacin in the treatment of a wide range of diseases in dogs and cats.     

Enhanced intraphagocytic killing of Brucella abortus in bovine mononuclear cells by liposomes-containing gentamicin

In vitro intraphagocytic killing of Brucella abortus in bovine mononuclear leukocytes was enhanced by cationic, anionic, and neutral multilamellar liposomes-containing gentamicin. Free gentamicin not entrapped in liposomes. and liposomes without antibiotic did not enhance intraphagocytic killing of B. abortus in bovine phagocytes. In vivo killing of B. abortus in guinea pigs was also enhanced by liposomes-containing gentamicin when compared to free gentamicin. Liposomes-containing alpha tocopherol acetate failed to enhance in vivo killing of B. abortus.     

In vitro activity of difloxacin against canine bacterial isolates.

The in vitro activity of difloxacin against canine bacterial isolates from clinical cases was studied in the United States and The Netherlands. Minimal inhibitory concentrations (MIC), the postantibiotic effect, the effect of pH on antimicrobial activity, and the bacterial killing rate tests were determined according to standard techniques. The MICs of American and Dutch isolates agreed in general. The MICs of the American gram-negative isolates ranged from 0.06 to 2.0 microg/ml, and the MICs of the Dutch gram-negative isolates ranged from 0.016 to 8.0 microg/ml. A few European strains of Proteus mirabilis and Klebsiella pneumoniae had relatively high MICs. Bordetella bronchiseptica also was less susceptible to difloxacin. The MICs of the American gram-positive cocci ranged from 0.125 to 4.0 microg/ml, and the MICs of Dutch isolates ranged from 0.125 to 2.0 microg/ ml. Difloxacin induced a concentration-dependent postantibiotic effect that lasted 0.2-3 hours in cultures with Escherichia coli, Staphylococcus intermedius, Streptococcus canis, Proteus spp., and Klebsiella pneumoniae. There was no postantibiotic effect observed against canine Pseudomonas aeruginosa. Decreasing the pH of the medium increased the MIC of Proteus mirabilis for difloxacin. The MICs of Escherichia coli and Klebsiella pneumoniae were lowest at neutral pH and were slightly increased in acid or alkaline media. At a neutral pH, most tested bacterial species were killed at a difloxacin concentration of 4 times the MIC. Similar results were obtained when these same bacteria were tested against enrofloxacin. A Klebsiella pneumoniae strain in an acidic environment was readily killed at difloxacin or enrofloxacin MIC, but at neutral pH the drug concentration had to be raised to 4 times the MIC for a bactericidal effect. After 24 hours of incubation at pH 7.1, difloxacin and enrofloxacin had similar bactericidal activity for all bacteria tested except Staphylococcus intermedius. Against S. intermedius, difloxacin was more bactericidal than enrofloxacin.     

Survival of Salmonella serovar Typhimurium inside porcine monocytes is associated with complement binding and suppression of the production of reactive oxygen species

The development of the carrier state in swine after infection with Salmonella serovar Typhimurium (S. Typhimurium) has not been elucidated yet. Possibly, phagocytes like macrophages play a crucial role. It was the aim of the present study to characterize the interaction of a S. Typhimurium strain and its hilA and ssrA mutants with porcine peripheral blood monocytes (PBM). Exposure of porcine PBM to S. Typhimurium induced the production of reactive oxygen species (ROS), requiring bacterial protein synthesis. The numbers of intracellular bacteria sharply decreased over a period of 3h. Monocytes obtained from different pigs differed markedly in their ROS production and in their ability to kill the bacteria. Interestingly, high ROS production did not coincide with increased intracellular killing. Using diphenylene iodonium inhibition of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity, it was shown that bacterial killing was ROS-dependent only within 1h post inoculation, but was ROS-independent from 1h post inoculation onwards. This might be explained by the finding that metabolically active Salmonella bacteria were capable of suppressing the respiratory burst activity in a SPI-1- and SPI-2-independent manner without causing measurable cell damage. Opsonization with complement did not alter the ROS production. Nevertheless, it increased intracellular survival of the bacteria. In conclusion, survival of S. Typhimurium inside porcine PBM is promoted by suppression of respiratory burst activity and complement binding.     

The selection in vitro of antibiotics with activity against intracellular S. aureus

The activity of a range of antibiotics on intracellular Staphylococcus aureus was examined using an in vitro system in which staphylococci survived within bovine neutrophils and extracellular organisms were killed by lysostaphin. Cloxacillin in the presence of lysostaphin caused a reduction in the number of viable intracellular S. aureus but cloxacillin alone failed to reduce such bacteria significantly. The cloxacillin appeared to sensitize the staphylococci to lysis by extracellular traces of lysostaphin following neutrophil disruption. Extracellular staphylococci which remained viable after exposure to cloxacillin in the absence of lysostaphin were subsequently killed more rapidly by neutrophils, but this was not found with bacteria exposed to cloxacillin while inside cells. Vancomycin with lysostaphin produced a similar but smaller sensitization effect but this antibiotic also appeared to increase survival of intracellular staphylococci when compared with controls, possibly by impeding neutrophil bactericidal mechanisms. The only other antibiotic to show significant intracellular killing was rifampicin, and in this case the action was independent of lysostaphin.     

The influence of extracellular and phagolysosomal pH changes on the bactericidal activity of bovine neutrophils against Staphylococcus aureus

The influence of the pH of suspending medium on bovine neutrophil (PMN) function was assessed in tests of phagocytosis and killing of Staphylococcus aureus. Intracellular killing was markedly inhibited by moderate extracellular acidification whereas phagocytosis was little affected, except at the lowest pH level (pH 5.0). The killing of S. aureus by extracts of isolated PMN lysosomal granules showed a similar pH dependence and was optimal at pH levels above neutrality. Survival of S. aureus within PMN from different cows varied significantly and the relative differences in PMN bactericidal efficiency were maintained at all pH levels. The acidification of extracellular medium during incubation which resulted from metabolic activity of the PMN themselves, increased with increasing ratios of bacteria:PMN and varied significantly among cows. Addition of methylamine (10 mM) to elevate phagolysosomal pH inhibited phagocytosis and had no effect on intracellular survival of S. aureus. However, a lower concentration (1.5 mM) did not affect phagocytosis, but reduced bacterial survival without altering the relative differences in efficiency of PMN from different cows. It is suggested that the acidity of the extracellular medium may both reflect and influence the pH changes occurring within PMN phagosomes and, thereby, modulate the efficiency of intracellular destruction of S. aureus.     

Pharmacokinetics and tissue distribution of enrofloxacin and its active metabolite ciprofloxacin in calves

The purpose of this study was to establish the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin in the plasma and interstitial fluid (ISF) following subcutaneous (s.c.) administration of enrofloxacin. Ultrafiltration probes were placed in the s.c. tissue, gluteal musculature, and pleural space of five calves. Each calf received 12.5 mg/kg of enrofloxacin. Plasma and ISF samples were collected for 48 h after drug administration and analyzed by high pressure liquid chromatography. Plasma protein binding of enrofloxacin and ciprofloxacin was measured using a microcentrifugation system. Tissue probes were well tolerated and reliably produced fluid from each site. The mean +/- SD plasma half-life was 6.8 +/- 1.2 and 7.3 +/- 1 h for enrofloxacin and ciprofloxacin, respectively. The combined (ciprofloxacin + enrofloxacin) peak plasma concentration (Cmax) was 1.52 microg/mL, and the combined area under the curve (AUC) was 25.33 microg/mL. The plasma free drug concentrations were 54% and 81% for enrofloxacin and ciprofloxacin, respectively, and free drug concentration in the tissue fluid was higher than in plasma. We concluded that Cmax/MIC and AUC/MIC ratios for free drug concentrations in plasma and ISF would meet suggested ratios for a targeted MIC of 0.06 microg/mL.     

恩诺沙星与硫酸丁胺卡那霉素对奶牛乳房炎常见病原菌的联合药敏试验

本试验对恩诺沙星和硫酸丁胺卡那霉素在4种奶牛乳房炎常见病原菌（无乳链球菌、化脓链球菌、金黄色葡萄球菌、致病性大肠埃希氏菌）联合药敏试验中的相互作用进行了研究。结果表明,恩诺沙星和硫酸丁胺卡那霉素对各菌的MIC为8～64μg／mL,二者联合后的FIC指数范围为0．5～1,二者联合用药时为协同作用或相加作用。     

In vitro killing of Escherichia coli, Staphylococcus pseudintermedius and Pseudomonas aeruginosa by enrofloxacin in combination with its active metabolite ciprofloxacin using clinically relevant drug concentrations in the dog and cat

Enrofloxacin is a fluoroquinolone antibacterial agent used to treat infections in companion animals. Enrofloxacin's antimicrobial spectrum includes Gram positive and Gram-negative bacteria and demonstrates concentration-dependent bacteriocidal activity. In dogs and cats, enrofloxacin is partially metabolized to ciprofloxacin and both active agents circulate simultaneously in treated animals at ratios of approximately 60-70% enrofloxacin to 30-40% ciprofloxacin. We were interested in determining the killing of companion animal isolates of Escherichia coli, Staphylococcus pseudintermedius and Pseudomonas aeruginosa by enrofloxacin and ciprofloxacin combined using clinically relevant drug concentrations and ratios. For E. coli isolates exposed to 2.1 and 4.1μg/ml of enrofloxacin/ciprofloxacin at 50:50, 60:40 and 70:30 ratios, a 1.7-2.5log(10) reduction (94-99% kill) was seen following 20min of drug exposure; 0.89-1.7log(10) (92-99% kill) of S. pseudintermedius following 180min of drug exposure; 0.85-3.4log(10) (98-99% kill) of P. aeruginosa following 15min of drug exposure. Killing of S. pseudintermedius was enhanced in the presence of enrofloxacin whereas killing of P. aeruginosa was enhanced in the presence of ciprofloxacin. Antagonism was not seen when enrofloxacin and ciprofloxacin were used in kill assays. The unique feature of partial metabolism of enrofloxacin to ciprofloxacin expands the spectrum of enhanced killing of common companion animal pathogens.     

Pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin in goats given enrofloxacin alone and in combination with probenecid.

The pharmacokinetics of enrofloxacin and its active metabolite ciprofloxacin were investigated in goats given enrofloxacin alone or in combination with probenecid. Enrofloxacin was administered i.m. at a dosage of 5 mg x kg(-1) alone or in conjunction with probenecid (40 mg x kg(-1), i.v.). Blood samples were drawn from the jugular vein at predetermined time intervals after drug injection. Plasma was separated and analysed simultaneously for enrofloxacin and ciprofloxacin by reverse-phase high performance liquid chromatography. The plasma concentration-time data for both enrofloxacin and ciprofloxacin were best described by a one-compartment open pharmacokinetic model. The elimination half-life (t(1/2beta)), area under the plasma concentration-time curve (AUC), volume of distribution (V(d(area))), mean residence time (MRT) and total systemic clearance (Cl(B)) were 1.39 h, 7.82 microg x h x mL, 1.52 L x kg(-1), 2.37 h and 802.9 mL x h(-1) x kg(-1), respectively. Enrofloxacin was metabolized to ciprofloxacin in goats and the ratio between the AUCs of ciprofloxacin and enrofloxacin was 0.34. The t(1/2beta), AUC and MRT of ciprofloxacin were 1.82 h, 2.55 microg x h x mL and 3.59 h, respectively. Following combined administration of probenecid and enrofloxacin in goats, the sum of concentrations of enrofloxacin and ciprofloxacin levels > or = 0.1 microg x mL(-1) persisted in plasma up to 12 h.Co-administration of probenecid did not affect the t(1/2beta), AUC, V(d (area)) and Cl(B) of enrofloxacin, whereas the values of t(1/2beta) (3.85 h), AUC (6.29 microg x h x mL), MRT (7.34 h) and metabolite ratio (0.86) of ciprofloxacin were significantly increased. The sum of both enrofloxacin and ciprofloxacin levels was > or = 0.1 microg x mL(-1) and was maintained in plasma up to 8 h in goats after i.m. administration of enrofloxacin alone. These data indicate that a 12 h dosing regime may be appropriate for use in goats.