The effect of temperature and salinity on the elimination of enrofloxacin in the Manila clam Ruditapes philippinarum

The effect of temperature and salinity on the elimination of enrofloxacin (EF) in Manila clams Ruditapes philippinarum was investigated. The clams, cultured under different temperatures and salinities (16 degrees C and 30 per thousand, 22 degrees C and 30 per thousand, or 22 degrees C and 20 per thousand), were exposed to EF at 5 microg/mL of water in a medicated bath. After a 24-h exposure, the concentration of EF in various tissues was measured by high-performance liquid chromatography and the elimination rate of EF in those tissues was investigated by regression analysis. After the treatment, the initial concentrations of EF among tissues were (in decreasing order) plasma > gill > visceral mass > foot > adductor muscle. In all tissues the elimination half-life (t1/2) of EF in the clams cultured at 22 degrees C and 20 per thousand and 16 degrees C and 30 per thousand were markedly longer than in those cultured at 22 degrees C and 30 per thousand, and the t1/2 at 16 degrees C and 30 per thousand was slightly longer than that at 22 degrees C and 20 per thousand. Slight differences were also observed in t1/2 values among various tissues. These data indicate that both temperature and salinity had significant effects on the elimination of EF in the Manila clams and that lower temperature or salinity could result in slower elimination.     

Pharmacokinetics of difloxacin in olive flounder Paralichthys olivaceus at two water temperatures

In this study, the pharmacokinetics profiles of difloxacin in the olive flounder (Paralichthys olivaceus) were investigated following intravenous and oral administration (10 mg/kg BW) at 14 and 22 °C water temperatures. Plasma and tissue samples (muscle, liver, and kidney) were analyzed using an HPLC method. The results showed that the plasma concentration–time data for difloxacin were described commendably by two‐compartment open model at the two water temperatures. The absorption half‐life (t_1/2ka) of difloxacin after oral administration were 2.08 and 1.10 h at 14 and 22 °C, respectively; whereas the elimination half‐life (t_1/2β) was 4.41 and 2.38 h, respectively. The muscle concentration of 1.35 ± 0.19 μg/g was observed at 9 h at 14 °C, and 2.11 ± 0.33 μg/g at 6 h at 22 °C, respectively. For liver, the peak concentration of difloxacin 2.43 ± 0.30 μg/g occurred at 6 h at 14 °C, which was lower than the 3.34 ± 0.24 μg/g peak that occurred at 4 h at 22 °C. The calculated bioavailability of difloxacin was 68.07% at 22 °C, which was higher than the 53.43% calculated for 14 °C. After intravenous administration, the t_1/2β were 4.79 and 2.81 h at 14 and 22 °C, respectively. The results indicate that the peak concentrations in muscle and liver at 14 °C are approximately half of those achieved at 22 °C. However, the C_max in kidney at 14 and 22 °C were similar. The V_d values were 1.20 and 1.75 L/kg at 14 and 22 °C, respectively. These data indicated that both temperature and drug administration had significant effects on the elimination of difloxacin, and lower temperature or oral administration resulted in lower elimination.     

Pharmacokinetics of florfenicol and its metabolite,florfenicol amine,in rice field eel (Monopterus albus) after a single‐dose intramuscular or oral administration

The pharmacokinetics of florfenicol (FF) and its metabolite, florfenicol amine (FFA), were studied in rice field eel (Monopterus albus) after a single dose (20 mg/kg) by intramuscular (i.m.) or oral gavage (p.o.) dose at 25 °C. The elimination half‐lives (t_1/2β), peak concentration of FF (C_max), and time to reach FF peak concentration (T_max) in plasma were estimated as 18.39 h, 10.83 μg/mL, and 7.00 h, respectively, after i.m. injection and 13.46 h, 8.37 μg/mL, and 5 h, respectively, after p.o. administration. The T_max values of FF in tissues (i.e., kidney, muscle, and liver) were larger for i.m. injection compared with those for p.o. administration. The t_1/2β had the following order kidney > muscle > liver for i.m. administrated and kidney > liver > muscle for p.o. administrated. The largest area under the concentration–time curve (AUC) was calculated to be 384.29 mg · h/kg after i.m. dosing, and the mean residence time (MRT) was 42.46 h by oral administration in kidney. FFA was also found in all tissues with a lower concentration than FF for both i.m. and p.o. administrations throughout the study. The elimination of FFA was slow with a t_1/2β between 18.19 and 47.80 h in plasma and tissues. The mean metabolic rate of FFA for i.m. and p.o. administrations was >23.30%.     

Distribution of the portunid crab Ovalipes punctatus (De Haan) in Algoa Bay and salinity and temperature tolerances of its zoeae

The three-spot swimming crab Ovalipes punctatus is commonly found in surf zones off sandy beaches in Algoa Bay. It is exclusively marine and occurs down to a depth of 38 m. Large crabs were found close inshore while smaller crabs were predominantly found in deeper water, suggesting that settlement mainly occurs offshore in water 25 – 38 m deep. Gravid females were found closer inshore in shallow water. The zoeae could not tolerate high temperatures (>27,5 °C) or low salinities (<15‰). The optimal temperatures and salinities for survival of the zoeae were 15–22 °C and 34–35‰ respectively.     

Pharmacokinetics and tissue distribution of enrofloxacin after single intramuscular injection in Pacific white shrimp

The pharmacokinetic properties and tissue distribution of enrofloxacin (EF) were investigated after single intramuscular (i.m.) dose of 10 mg/kg body weight (b.w.) in Pacific white shrimp at 22 to 25°C. EF and its metabolite ciprofloxacin (CF) were determined by high‐performance liquid chromatography. After i.m. administration, EF was absorbed quickly, and the peak of EF concentration (C_max) reached at first time point in hemolymph. The volume of distribution V_d(area) of EF was 3.84 L/kg, indicating that the distribution of EF was good. The area under the concentration–time curve (AUC) of EF was 90.1 and 274.2 μg hr/ml in muscle and hepatopancreas, respectively, which was higher than 75.8 μg hr/ml in hemolymph. The EF elimination was slow in muscle and hepatopancreas with the half‐life (T_1/2β) of 52.3 and 75.8 hr, respectively. CF, the mainly metabolite of EF, was detected in hemolymph, muscle and hepatopancreas. The C_max was 0.030, 0.013 and 0.218 μg/ml, respectively. Based on a minimum inhibitory concentration (MIC) of 0.006–0.032 μg/ml for susceptible strains, EF i.m. injected at a dose 10 mg/kg could be efficacious against common pathogenic bacteria of Pacific white shrimp.     

Pharmacokinetics and tissue residues of moroxydine hydrochloride in gibel carp,Carassius gibelio after oral administration

The pharmacokinetics and tissue residues of moroxydine hydrochloride were studied in gibel carp at water temperature of 15 and 25 °C. Samples (blood, skin, muscle, liver, and kidney) were collected over 10 days after the treatment and analyzed by high‐performance liquid chromatography with an ultraviolet detector. The results indicated that the influence of water temperature on the metabolism of the drug was significant. The plasma concentration–time data of moroxydine hydrochloride conformed to single‐compartment open model at the two water temperatures. There were higher absorption rate (t_1/2ka) and longer elimination half‐lives (t_1/2ke) at 15 °C (4.29 and 15.87 h, respectively) compared with those at 25 °C (3.02 and 4.22 h, respectively). The maximum plasma concentration (C_max) and the time‐point of maximum plasma concentration (T_p) were 2.98 μg/mL and 10.35 h at 15 °C and 3.12 μg/mL and 4.03 h at 25 °C, respectively. The distribution volume (V_d/F) of moroxydine hydrochloride was estimated to be 4.55 L/kg at 15 °C and 2.89 L/kg at 25 °C. The total body clearance (CL_b) of moroxydine hydrochloride was determined to be 0.25 and 0.49 L/(h·kg) at 15 °C and 25 °C, respectively; the areas under the concentration–time curve were 75.89 μg·h/mL at 15 °C and 42.33 μg·h/mL at 25 °C. The depletion of moroxydine hydrochloride in gibel carp was slower with a longer half‐life period, especially at lower water temperature that was tested.     

Comparative pharmacokinetics and tissue distribution of quinocetone in crucian carp (Carassius auratus), common carp (Cyprinus carpio L.), and grass carp (Ctenopharyngodon idella) following the same experimental conditions

The pharmacokinetics and tissue distribution of quinocetone (QCT) in crucian carp (Carassius auratus), common carp (Cyprinus carpio L.), and grass carp (Ctenopharyngodon idella) were compared after oral administration of QCT (50 mg/kg body weight) at water temperature of 24 ± 1 °C. Similar QCT plasma concentration–time profiles were found in the three species of cyprinid fish at the same dosage regimen and water temperature, which were all fitted two‐compartment open pharmacokinetic model. However, different pharmacokinetic parameters were observed in crucian carp, common carp, and grass carp. The absorption rate constants (K_a) of QCT were 1.65, 1.40 and 1.74/h, respectively and absorption half‐lives (t_1/2kα) were 0.42, 0.49, and 0.40/h, respectively. The distribution half‐life (t_1/2α) was 2.83, 0.67, and 0.88 h, respectively, and elimination half‐lives (t_1/2β) of QCT were 133.97, 63.55, and 40.76 h, respectively. The maximum concentrations (C_max) of QCT in plasma were 0.315, 0.182, and 0.139 μg/mL and the time to peak concentrations (T_p) were 1.45, 0.96, and 1.08 h, respectively. The area under the plasma concentration‐time curves (AUC) were 12.35, 5.99, and 4.52 μg·h/mL, respectively. The distribution volumes (V_d/F) of QCT were calculated as 117.81, 128.71, and 220.10 L/kg, respectively. The tissue analysis showed that a similar regularity was obtained in the three species of cyprinids with a single dose of 50 mg/kg body weight after oral administration at the same water temperature. The tissue concentration of QCT in each fish was in order of liver>kidney>muscle, while the residues of QCT in the three species of cyprinid fish were in order of crucian carp>common carp>grass carp.     

Pharmacokinetics of florfenicol in blunt‐snout bream (Megalobrama amblycephala) at two water temperatures with single‐dose oral administration

The pharmacokinetics and residue elimination of florfenicol (FFC) and its metabolite florfenicol amine (FFA) were studied in healthy blunt‐snout bream (Megalobrama amblycephala, 50 ± 10 g). The study was conducted with a single‐dose (25 mg/kg) oral administration at a water temperature of 18 or 28°C, while in the residue elimination study, fish were administered at 25 mg/kg daily for three consecutive days by oral gavage to determine the withdrawal period (WDT) at 28°C. The FFC and FFA levels in plasma and tissues (liver, kidneys and muscle) were analysed using high‐performance liquid chromatography (HPLC). A no‐compartment model was used to analyse the concentration versus time data of M. amblycephala. In the two groups at 18 and 28°C, the maximum plasma concentration (C_max) of FFC was 5.89 and 6.21 μg/ml, while the time to reach C_max (T_max) was 5.97 and 2.84 hr, respectively. These suggested that higher temperature absorbed more drug and more quickly at M. amblycephala. And the elimination half‐life (T_1/2kβ) of FFC was calculated as 26.75 and 16.14 hr, while the total body clearance (CL) was 0.09 and 0.15 L kg^?1 hr^?1, and the areas under the concentration–time curves (AUCs) were 265.87 and 163.31 μg hr/ml, respectively. The difference demonstrated that the elimination rate of FFC in M. amblycephala at 28°C was more quickly than that at 18°C. The results of FFA showed the same trend in tissues of M. amblycephala. After multiple oral doses (25 mg/kg daily for 3 days), the k (eliminate rate constant) of FFA in M. amblycephala muscle was 0.017, the C₀ (initial concentration) was 3.07 mg/kg, and the WDT was 10 days (water temperature 28°C).     

Pharmacokinetics of florfenicol and its metabolite florfenicol amine in crucian carp (Carassius auratus) at three temperatures after one single intramuscular injection

The pharmacokinetic profiles of florfenicol (FF) or florfenicol amine (FFA) in crucian carp were compared at different water temperatures after single intramuscular administration of FF at 10 mg/kg bodyweight. The concentrations of FF and FFA were determined by a high‐performance liquid chromatography method, and then, the concentration versus time data were subjected to compartmental analysis using a one‐compartment open model. At the water temperatures of 10, 20, and 25°C, the peak concentrations (C_maxs) of FF were 2.28, 2.29, and 2.34 μg/ml, respectively, while those of FFA were 0.42, 0.71, and 0.82 μg/ml, respectively. And the absorption half‐life (t_1/2ka) of FF was 0.21, 0.19, and 0.21 hr, while the elimination half‐life (t_1/2kel) was 31.66, 24.77, and 21.48 hr, respectively. For FFA, the formation half‐life (t_1/2kf) was 3.85, 8.97, and 12.43 hr, while the t_1/2kel was 58.34, 30.27, and 21.22 hr, respectively. The results presented here demonstrated that the water temperature had effects on the elimination of both FF and FFA and the formation of FFA. Based on the T > MIC values calculated here, to treat the infections of bacterial with MIC value ≤ 0.5 μg/ml, FF intramuscularly given at 10 mg/kg bodyweight with a 72‐hr interval is sufficient at the water temperature of 10°C, while the intervals of 60 and 48 hr were needed at 20 and 25°C, respectively. But to treat bacterial with higher MIC values, more FF or FF at 10 mg/kg BW but with shorter intervals should be intramuscularly given to the infected fish.     

Pharmacokinetics of enrofloxacin in snakehead fish,Channa argus

The pharmacokinetics of enrofloxacin (EF) was investigated after single intravenous (i.v.) and oral (p.o.) dose of 10 mg/kg body weight (b.w.) in snakehead fish at 24–26 °C. The plasma concentrations of EF and its metabolite ciprofloxacin (CF) were determined by high‐performance liquid chromatography. The plasma concentration–time data were described by an open two‐compartment model for both routes. After intravenous administration, the elimination half‐life (T_1/2β), area under the concentration–time curve (AUC) and total body clearance of EF were 19.82 h, 75.79 μg h/mL and 0.13 L/h/kg, respectively. Following p.o. administration, the maximum plasma concentration (C_max), T_1/2β and AUC of EF were 1.86 μg/mL, 35.8 h and 49.98 μg h/mL, respectively. Absorption of EF was good with a bioavailability (F) of 65.82%, which was higher than that calculated in most seawater fish. CF, an active metabolite of EF, was detected occasionally in this study, which indicates a low extent of deethylation of EF in snakehead fish.     

Pharmacokinetic profiles of florfenicol in spotted halibut,Verasper variegatus,at two water temperatures

The pharmacokinetic profiles of florfenicol in the spotted halibut (Verasper variegatus) were investigated at 15 and 20°C water temperatures, respectively. Florfenicol content in plasma samples was analyzed using an HPLC method. Drug concentration versus time data were best fitted to a three‐compartment model after a single intravenous administration (15 mg/kg BW), and fitted to a two‐compartment model after an oral administration (30 mg/kg BW) at 15 and 20°C. The florfenicol concentration in the blood increased slowly during the 12 hr following an oral administration at 15°C, with a peak concentration (C_max) of 9.1 mg/L, and then declined gradually. The half‐lives of absorption, distribution, and elimination phase were 2.18, 5.66 and 14.25 hr, respectively. The bioavailability (F) was calculated to be 24.14%. After an oral administration at 20°C, shorter half‐lives of absorption (1.33 hr), distribution (2.51 hr) and elimination (9.71 hr), a higher C_max (12.2 mg/L), and a similar F (23.98%) were found. Based on the pharmacokinetics and pharmacodynamics, an oral dose of 30 mg/kg BW was suggested to be efficacious for bacterial disease control in spotted halibut farming.     

Tilmicosin antibacterial activity and pharmacokinetics in cows

The minimal inhibitory concentration (MIC) of tilmicosin for 90% of 112 Staphylococcus aureus isolates from the bovine udder was 0.78 μg/mL and 149 of 164 (90.8%) other gram-positive udder pathogens were inhibited by tilmicosin concentrations < 3.12 μg/mL. The MIC of the drug for 19 of 22 S. aureus isolates was < 0.78 μg/mL when the test was conducted using Mueller-Hinton (MH) agar or MH agar containing 7.5% skimmed milk. Acute cardiac toxicity followed intravenous (i.v.) injection of the drug at 10 mg/kg to 3 cows, but animals appeared clinically normal within 30 min after treatment. The pharmacokinetics of i.v.-administered tilmicosin is typical for the macrolide class of antibiotics, i.e. low serum drug concentrations and a large volume of distribution (> 2.0 L/kg). The elimination half-life (t_1/2β values for 3 cows were 46.4. 56.0 and 72.8 min. The drug was administered subcutaneously (s.c.) to 5 cows at 10 mg/kg; the elimination half-life (t_1/2el) was 4.18 ± 0.55 h and the mean s.c. bioavailability was 22%. Rapid and extensive penetration of tilmicosin from blood into milk, and slow elimination from the milk were among the characteristic kinetic features of the drug after i.v. and s.c. administration. Tilmicosin was injected s.c. at 10 mg/kg once to 9 cows after the last milking of lactation; dry udder secretion samples were collected daily for 11 consecutive days and assayed microbiologically. Concentrations of drug > 0.78 μg/mL were found in the secretion for 8–9 days after dosing. Systemic side-effects were not observed after s.c. drug administration.     

The effects of wilting and storage temperatures on the fermentation quality and aerobic stability of stylo silage

In order to clarify the ensiling characteristics of stylo (Stylosanthes guianensis Swartz), the effects of wilting (no wilting, light wilting and heavy wilting) and storage temperatures (10°C, 20°C, 30°C and 40°C) on the fermentation quality and aerobic stability of stylo silage were investigated. Wilting had no significant influence on the contents of crude protein, ether extract and acid detergent fiber, and numbers of lactic acid bacteria, aerobic bacteria, yeasts and mold (P > 0.05). Heavy wilted material, wilted for 12 h, had higher neutral detergent fiber content and lower water‐soluble carbohydrate content than unwilted and light wilted materials (P < 0.05). Wilting and storage temperatures had significant effects on pH value, acetic acid, butyric acid and NH₃‐N contents of stylo silage (P < 0.01 or P < 0.05). Wilting tended to reduce acetic acid and NH₃‐N contents and improve the fermentation quality of stylo silage. In all the silages, no wilting silage ensiled at 30°C had the highest butyric acid content (P < 0.05). High temperature of 40°C markedly restricted the growth of lactic acid bacteria and aerobic bacteria in silage, irrespective of wilting. The wilted silage or silage stored at low temperature had poor aerobic stability.     

The effect of temperature and salinity on oxygen consumption in the brachyuran crab Cyclograpsus punctatus (Crustacea: Decapoda: Grapsidae)

The aquatic oxygen consumption of the estuarine brachyuran crab, Cyclograpsus punctatus, was investigated after a 24-hour acclimation period at different temperature (12.5, 20, 30°C) and salinity (9, 17.5, 35, and 44°) combinations . Salinity had no significant effect on oxygen consumption at 12.5 and 20°C in both large and small crabs. At 30°C and 44°, however, the rate of oxygen consumption declined in large crabs (from 0.233 to 0.176 mg O₂ g wwt/h) and increased in small crabs (0.300 to 0.469 mg O₂ g wwt/h). There was also a significant increase in the oxygen consumption (0.300 to 0.536 mg O₂ g wwt/h) of small crabs at the high temperature/low salinity combination. Temperature had a significant effect on the rate of oxygen consumption at all salinities, with Q₁₀ values ranging from 1.16 to 2.85. In all cases the mass-specific oxygen consumption rate of small crabs was higher than that of large crabs. The results suggest that the confinement of this species to the lower reaches of estuaries may, in part, be due to physiological constraints on juveniles and small individuals.     

Effect of Incubation Temperature and Salinity on Expression of the Outer Membrane Protein Profile of Edwardsiella tarda

Abstract

Outer membrane proteins (OMP) of 10 isolates of Edwardsiella tarda were compared by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis. The OMP profile of the type strain E. tarda ATCC 15947 cultured at 25°C had five major protein bands of 40, 36.5, 34, 28.5, and 25 kDa and a large number of minor proteins ranging in size from approximately 10 to 120 kDa. Differences between the OMP profiles of the isolates of E. tarda included the inconsistent presence of the 34- or 36.5-kDa proteins in five isolates of E. tarda and two major bands of 47 and 44 kDa that were present in only two isolates of E. tarda. There were no differences in the outer membrane protein profiles of 9 out of 10 isolates of E. tarda incubated at a temperature of 25°C compared with those at 35°C. To evaluate the effect of salinity, 10 isolates of E. tarda were cultured in brain heart infusion broth containing 0.5, 1.5, and 3.0% sodium chloride. Reactions of isolates of E. tarda to the different salinity levels were placed into three groups. The first group expressed more or fewer protein bands at 1.5% sodium chloride. The second group lost major bands at 3% salinity, whereas the third group had no change in the OMP profile with salinity. The OMP profile differences and the different reactions to salinity levels suggest that the isolates are heterogeneous.     

Pharmacokinetic Profile of Erythromycin after Intramammary Administration in Lactating Dairy Cows with Specific Mastitis

The pharmacokinetics of erythromycin was studied in five lactating dairy cows following single intramammary infusion of 300 mg erythromycin in each of two quarters per cow with specific mastitis. Levels of erythromycin in plasma and quarter milk samples were measured by agar plate diffusion assay using Micrococcus luteus (ATCC 9341) as the test organism. Erythromycin level in plasma reached a peak concentration value (C _max) of 0.07 ± 0.01 μg/ml at 30 min; thereafter, levels declined gradually to reach 0.05 ± 0.00 μg/ml 12 h post drug administration. The pharmacokinetic profile of the drug revealed mean absorption half life (t _1/2ka) as 0.26 ± 0.05 h. The drug was eliminated slowly with elimination half-life (t _1/2β) of 13.75 ± 0.35 h and elimination rate constant (k _el) of 0.04 ± 0.00 h⁻¹. The volume of distribution based on the zero-time plasma concentration intercept of the least-squares regression line of the elimination phase (V _d(B)) was 0.032 L/kg. The drug crossed to untreated quarters also; mean drug levels of 0.20 ± 0.07, 0.23 ± 0.07, 0.17 ± 0.04, and 0.17 ± 0.04 μg/ml were found at 3, 6, 8 and 12 h, respectively. The mean drug concentration for treated quarters was measured as 22.97 ± 2.31 μg/ml milk at first milking (12 h) following drug infusion. No apparent adverse reaction was seen in cows administered erythromycin. It is concluded that following intramammary infusion erythromycin diffuses readily and extensively in various body fluids and tissues and adequate concentration is maintained in udder tissues for at least 12 h post intramammary administration. Thus, erythromycin may be recommended for local therapy of acute mastitis caused by Gram-positive bacteria in lactating dairy cows.     

Ultrastructural Alterations in the Gills of Labeo rohita Fingerlings Exposed to Thermal Extremes

This study aimed to determine the cellular alterations in the gill of Labeo rohita exposed to lethal temperature maxima (LT_Max) and lethal temperature minima (LT_Min) by means of transmission electron microscopy (TEM). Acclimation of advanced fingerlings of L. rohita was carried out at 26°C for 30 days. Acclimated fish were subjected to a constant rate of increase or decrease in temperature (0.3°C/min) until the LT_Max and LT_Min values were reached. Dissected gills were processed for TEM, both at the end of acclimation period at ambient temperature (26°C) and at lethal temperatures. Results indicated that at ambient temperature, the gill tissues appeared normal. However, significant changes were observed at lethal temperatures. The gill tissues at lethal temperature maxima showed severely damaged lamellae, with more vacuolated space. At lethal temperature minima, gill tissues showed increased density of mitochondria. Our prima‐facie report indicated that L. rohita exposed to lethal temperatures exhibited marked ultrastructural changes in the gills.     

A Comparison of Five Cooling Methods in Hot and Humid Environments in Thoroughbred Horses

Horses need aggressive cooling to prevent exertional heat illness after strenuous exercise in hot and humid conditions. This study compared various methods for cooling horses in such conditions, testing the hypothesis that continual application of running water would be the most effective method to decrease core temperature. Five Thoroughbreds were exercised on a treadmill at wet-bulb globe temperature of 31.8 ± 0.1°C until their pulmonary artery temperature reached 42°C. The time until the pulmonary artery temperature returned to <39°C (t₃₉) and the rectal temperature at 30 minutes after the onset of cooling were compared between five cooling methods in a 5 × 5 Latin square design: walking, with no additional cooling (CONT); walking, with fans producing an air current of 3.0 m/s (FAN); walking, with the intermittent application of cold water (10°C) either with scraping (ICW + SCRAPE) or without scraping (ICW); and stationary, with the continuous tap water (26°C) application via shower hoses (STW). The STW produced the shortest t₃₉ (P < .001). With intermittent cold water, ICW + SCRAPE tended to produce longer t₃₉ than ICW (P = .06), and both produced significantly shorter t₃₉ than FAN and CONT (P < .001). The t₃₉ of the FAN tended to be shorter than with the CONT (P = .06). All the cooling methods resulted in lower rectal temperatures at 30 minutes than CONT. The temperature was the lowest with the shower method, with no significant differences between the fan and intermittent cold-water methods. Showering with tap water was the most effective method to decrease core temperature in horses.     

Pharmacokinetics of doxycycline in tilapia (Oreochromis aureus × Oreochromis niloticus) after intravenous and oral administration

The pharmacokinetics of doxycycline was studied in plasma after a single dose (20 mg/kg) of intravenous or oral administration to tilapia (Oreochromis aureus × Oreochromis niloticus) reared in fresh water at 24 °C. Plasma samples were collected from six fish per sampling point. Doxycycline concentrations were determined by high‐performance liquid chromatography with a 0.005 μg/mL limit of detection, then were subjected to noncompartmental analysis. Following oral administration, the double‐peak phenomenon was observed, and the first (C_max1) and second (C_max2) peaks were 1.99 ± 0.43 μg/mL at 2.0 h and 2.27 ± 0.38 μg/mL at 24.0 h, respectively. After the intravenous injection, a C_max2 (12.12 ± 1.97 μg/mL) was also observed, and initial concentration of 45.76 μg/mL, apparent elimination rate constant (λ_z) of 0.018 per h, apparent elimination half‐life (t_1/2λz) of 39.0 h, systemic total body clearance (Cl) of 41.28 mL/h/kg, volume of distribution (Vz) of 2323.21 mL/kg, and volume of distribution at steady‐state (Vss) of 1356.69 mL/kg were determined, respectively. While after oral administration, the λ_z, t_1/2λz, and bioavailability of doxycycline were 0.009 per h, 77.2 h, and 23.41%, respectively. It was shown that doxycycline was relatively slowly and incompletely absorbed, extensively distributed, and slowly eliminated in tilapia, in addition, doxycycline might undergo enterohepatic recycling in tilapia.     

In vitro production of porcine zygotes using intracytoplasmic injection of vitrified sperm

The objectives of this study were to evaluate if vitrified porcine spermatozoa are able to maintain their capacity to produce zygotes in vitro using intracytoplasmic sperm injection (ICSI) and to evaluate the zygote development in two in vitro atmospheric conditions: 5% CO₂ and tri‐gas. A group of porcine oocytes maturated in vitro were injected with vitrified‐warmed sperm (treatment group) and another group, with sperm diluted and conserved at 17°C (control group). To evidence parthenogenetic activation, some oocytes were submitted to a Sham test. The injected oocytes were cultured in G1 medium at 38°C, 100% humidity and 5% CO₂ or tri‐gas. No significant differences (p > .05) were observed in embryo development between the oocytes injected with vitrified‐warmed sperm (31.8%; 36/113), and those injected with semen diluted and conserved at 17°C (35.5%; 32/90), when cultured in 5% CO₂ or under tri‐gas atmosphere (42.9%; 39/91 vs. 34.2%; 26/76, respectively). No significant differences (p > .05) were observed in the percentage of pronuclei (PN) obtained between 5% CO₂ and tri‐gas, within each treatment either. Of the 52 oocytes submitted to the Sham test, only two presented a female PN (activation) indicating that the PN observed in the treatment group were a product of fertilization and not parthenogenetic activation. To conclude, porcine sperm vitrified using spheres, at a concentration of 5 × 10⁶ spermatozoa/ml in TALP medium with 1% bovine serum albumin (BSA), conserve condensed and intact chromatin capable of producing early embryo development up to the pronuclear stage.