Ontogeny expression of ghrelin,neuropeptide Y and cholecystokinin in blunt snout bream,Megalobrama amblycephala

Ghrelin, neuropeptide Y (NPY) and cholecystokinin (CCK) all have important roles in the regulation of feeding in fish and mammals. To better understand the role of the three peptides in appetite regulation in the early developmental stages of blunt snout bream (Megalobrama amblycephala), partial cDNA sequences of ghrelin, NPY and CCK genes were cloned. And then, real‐time quantitative PCR and RT‐PCR were used to detect and quantify the mRNA expressions of these genes from zygotes to larvae of 50 days after hatching (DAH). Ghrelin, NPY and CCK were all expressed throughout the embryonic and larval development stages, and the expression levels were higher in larval stages than in embryonic stages. Ghrelin and NPY mRNA expressions were upregulated at 1, 3, 5 DAH, while CCK mRNA expression was reduced significantly at 3 DAH. The mRNA expression levels of three genes in larvae varied significantly until 30 DAH. In adult fish, all three peptides were detected to be expressed in brain and several peripheral tissues. Ghrelin mRNA was mainly expressed in the intestine, whereas NPY and CCK mRNAs were mainly expressed in the brain. Taken together, these results indicate that ghrelin, NPY and CCK may have roles in early development and participate in the regulation of feeding of larvae in blunt snout bream and will be helpful for further investigation into feed intake regulation in adults of this species.     

幼鱼和育成鱼阶段团头鲂对饲料中脂肪的需要量

本试验旨在研究相同饲料配方体系下2个生长阶段——幼鱼和育成鱼阶段团头鲂对饲料中脂肪的需要量。以进口鱼粉、酪蛋白为蛋白质源,以大豆卵磷脂、豆油为脂肪源,在等蛋白质(粗蛋白质含量为30%)的条件下,配制脂肪水平(实测值)分别为3.06%、4.38%、5.89%、7.45%、8.78%、10.13%的6种半纯化饲料。幼鱼和育成鱼阶段试验分别以初始均重为(34.98±0.47)g/尾、(102.05±2.31)g/尾的团头鲂为试验鱼,以不同脂肪水平的试验饲料在池塘网箱中分别养殖85、56 d。同一生长阶段下,每种饲料为1组,每组4个重复,每个重复20尾鱼。结果表明:随着饲料脂肪水平的升高,2个生长阶段团头鲂的增重率、蛋白质效率均呈现先上升后下降的趋势,并均在饲料脂肪水平为8.78%时达到最高值;饲料系数则呈现先下降后上升的趋势,且均在饲料脂肪水平为8.78%时达到最低值。在相同的饲料配方体系下,团头鲂全鱼粗蛋白质含量表现为幼鱼阶段<育成鱼阶段,粗脂肪含量则表现为育成鱼阶段>幼鱼阶段。随着饲料脂肪水平的升高,2个生长阶段团头鲂全鱼粗蛋白质含量总体均呈现先上升后下降的趋势;粗脂肪含量则随着饲料脂肪水平的升高而增加,显示出与饲料脂肪水平具有正相关关系。以增重率、饲料系数、蛋白质效率作为评价指标,经过回归分析可知,幼鱼阶段团头鲂对饲料中脂肪的需要量为8.18%~8.89%,育成鱼阶段团头鲂对饲料中脂肪的需要量为9.03%~9.47%。     

氟苯尼考纳米晶在鸡体内的药动学

氟苯尼考(florfenicol, FFC)是新型动物广谱抗菌剂,抗菌效果好,广泛应用于牛、羊、猪、水产及禽类等动物细菌性疾病的防制。本试验旨在研究FFC和氟苯尼考纳米晶(florfenicol nanocrystal, FFC-NC)在鸡体内的生物利用度。采用交叉试验法,鸡用药后,在不同时间点翅下静脉采血,利用高效液相色谱法(high performance liquid chromatography, HPLC)测定血浆中FFC含量。结果显示,此试验所建立的HPLC色谱图基线平稳,血浆峰与FFC峰完全分离。回收率和精密度均符合测定要求,重复性好,适用于鸡血浆FFC含量测定。药动学参数结果显示,与FFC组相比,FFC-NC组的达峰时间t_max为(0.875±0.137) h,峰时缩短,药时曲线下面积AUC₍(0-∞))和峰浓度C_max分别为(23.957±2.338) mg/(L·h)和(8.249±0.713) mg/L,FFC-NC组的相对生物利用度是FFC组的3.6倍。结果表明,FFC-NC的药动学特征较FFC均...     

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 and bioavailability of flumequine in blunt snout bream (Megalobrama amblycephala) after intravascular and oral administrations

In this study, the pharmacokinetic profile of flumequine (FMQ) was investigated in blunt snout bream (Megalobrama amblycephala) after intravascular (3 mg/kg body weight (b.w.)) and oral (50 mg/kg b.w.) administrations. The plasma samples were determinedby ultra‐performance liquid chromatography (UPLC) with fluorescence detection. After intravascular administration, plasma concentration–time curves were best described by a two‐compartment open model. The distribution half‐life (t_1/2α), elimination half‐life (t_1/2β), and area under the concentration–time curve (AUC) of blunt snout bream were 0.6 h, 25.0 h, and 10612.7 h·μg/L, respectively. After oral administration, a two‐compartment open model with first‐order absorption was also best fit the data of plasma. The t_1/2α, t_1/2β, peak concentration (C_max), time‐to‐peak concentration (T_max), and AUC of blunt snout bream were estimated to be 2.5 h, 19.7 h, 3946.5 μg/L, 1.4 h, and 56618.1 h. μg/L, respectively. The oral bioavailability (F) was 32.0%. The pharmacokinetics of FMQ in blunt snout bream displayed low bioavailability, rapid absorption, and rapid 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%.     

Pharmacokinetics and dynamics of mycophenolate mofetil after single‐dose oral administration in juvenile dachshunds

Mycophenolate mofetil (MMF) is recommended as an alternative/complementary immunosuppressant. Pharmacokinetic and dynamic effects of MMF are unknown in young‐aged dogs. We investigated the pharmacokinetics and pharmacodynamics of single oral dose MMF metabolite, mycophenolic acid (MPA), in healthy juvenile dogs purpose‐bred for the tripeptidyl peptidase 1 gene (TPP1) mutation. The dogs were heterozygous for the mutation (nonaffected carriers). Six dogs received 13 mg/kg oral MMF and two placebo. Pharmacokinetic parameters derived from plasma MPA were evaluated. Whole‐blood mitogen‐stimulated T‐cell proliferation was determined using a flow cytometric assay. Plasma MPA C_max (mean ± SD, 9.33 ± 7.04 μg/ml) occurred at <1 hr. The AUC_0–∞ (mean ± SD, 12.84±6.62 hr*μg/ml), MRT_inf (mean ± SD, 11.09 ± 9.63 min), T1/2 (harmonic mean ± PseudoSD 5.50 ± 3.80 min), and k/d (mean ± SD, 0.002 ± 0.001 1/min). Significant differences could not be detected between % inhibition of proliferating CD5+ T lymphocytes at any time point (p = .380). No relationship was observed between MPA concentration and % inhibition of proliferating CD5+ T lymphocytes (R = .148, p = .324). Pharmacodynamics do not support the use of MMF in juvenile dogs at the administered dose based on existing therapeutic targets.     

Comparative single‐dose pharmacokinetics of orally administered florfenicol in rainbow trout (Oncorhynchus mykiss,Walbaum, 1792) at health and experimental infection with Streptococcus iniae or Lactococcus garvieae

This study evaluates changes in the pharmacokinetic behavior of a single oral dose of florfenicol in rainbow trouts experimentally infected with Lactococcus garvieae or Streptococcus iniae. One hundred and fifty fish were randomly divided into three equal groups: 1—healthy fish, 2—fish inoculated with S. iniae (2.87 × 10⁷ CFU/ml, i.p.), and 3—fish inoculated with L. garvieae (6.8 × 10⁵ CFU/ml, i.p.). Florfenicol was administered to all groups at 15 mg/kg by oral gavage. Blood sampling was performed at 0, 2, 3, 6, 8, 12, 24, 48, 72, and 120 hr after drug administration to each group, and plasma concentration of florfenicol was assayed by HPLC method. The MICs of florfenicol were 1.2 μg/ml and 5 μg/ml against L. garviae and S. iniae, respectively. Healthy fish showed higher values for most of the PK/PD parameters as compared to fish infected with L. garvieae which was reversed in fish infected with S. iniae. Fish infected with L. garvieae showed decreased relative bioavailability accompanied by increased volume of distribution at steady‐state (Vd_ss) and total body clearance (Cl_B)_. Infection with S. iniae increased the peak concentration of drug after administration (Cmax) and decreased elimination half‐life (T_{1/2 β})_, central compartment volume (V_c), and Vd_ss. In conclusion, infection with these bacteria can affect the pharmacokinetic behavior of florfenicol in rainbow trouts as shown by decreased bioavailability and increased total body clearance and volume of distribution in L. garvieae infection and decreased volume of distribution accompanied by increased Cmax in S. iniae‐infected fish.     

Pharmacokinetics of levosulpiride after single‐dose administration in goats (Capra hircus) by different routes of administration

Levosulpiride (LSP) is the l‐enantiomer of sulpiride, and LSP recently replacing sulpiride in several EU countries. Several studies about LSP in humans are present in the literature, but neither pharmacodynamic nor pharmacokinetic data of LSP is present for veterinary species. The aim of this study was to assess the pharmacokinetic profile of LSP after intravenous (IV), intramuscular (IM), and oral (PO) administration in goats. Animals (n = 6) were treated with 50 mg LSP by IV, IM, and PO routes according to a randomized cross‐over design (3 × 3 Latin‐square). Blood samples were collected prior and up to 24 hr after LSP administration and quantified using a validated HPLC method with fluorescence detection. IV and IM administration gave similar concentration versus time curve profiles. The IM mean bioavailability was 66.97%. After PO administration, the drug plasma concentrations were detectable only in the time range 1.5–4 hr, and the bioavailability (4.73%) was low. When the AUC was related to the administered dose in mg/kg, there was a good correlation in the IV and IM groups, but very low correlation for the PO route. In conclusion, the IM and IV administrations result in very similar plasma concentrations. Oral dosing of LSP in goats is probably not viable as its oral bioavailability was very low.     

高温季节饥饿后补偿摄食对团头鲂幼鱼生长性能、血清生化指标、肠道消化酶活性及肝脏抗氧化酶活性的影响

本试验旨在探讨高温季节饥饿后补偿摄食对团头鲂幼鱼生长性能、血清生化指标、肠道消化酶活性及肝脏抗氧化酶活性的影响。试验设计5个组,分别为饥饿0、5、10、15、20 d后再补偿摄食3周,依次标记为S0(对照)、S5、S10、S15、S20组。选择初始均重为(22.03±0.04)g的团头鲂幼鱼270尾,随机分入这5组,每组3个重复,每个重复18尾。结果显示:1)S10组试验鱼末体均重、增重率、特定生长率和饲料效率均达到最高,且补偿摄食期间其日摄食量和饲料效率显著高于对照组(P0.05),说明团头鲂幼鱼补偿生长是通过提高摄食率和饲料效率共同实现的。2)S5、S10组血清甘油三酯水平显著低于其他组(P0.05),而S20组血清谷草转氨酶活性显著高于其他组(P0.05),说明饥饿时间过长鱼体肝脏受到一定程度损伤;血清中其他生化指标如葡萄糖、总蛋白、白蛋白、总胆固醇水平和谷丙转氨酶活性在各组间无显著差异(P0.05)。3)补偿摄食后,S15组肠道脂肪酶活性显著高于其他组(P0.05);S20组肠道蛋白酶活性显著高于其他组(除S5组外)(P0.05),说明饥饿时间过长在补偿摄食后鱼体内蛋白质消化能力增强。4)各饥饿后补偿摄食组肝脏超氧化物歧化酶(除S5、S20组外)和过氧化氢酶活性均显著高于对照组(P0.05),而S20组肝脏谷胱甘肽过氧化物酶活性显著低于其他饥饿后补偿摄食组(P0.05),说明饥饿时间过长对机体抗氧化能力的恢复有所影响。由此可见,在本试验中,高温季节团头鲂幼鱼饥饿10 d后补偿摄食3周,补偿生长效果达到最佳,鱼体免疫与抗氧化能力也得到一定增强。     

液-液-液微萃取与高效液相色谱联用技术测定饲料中的己烯雌酚

建立了液-液-液微萃取与高效液相色谱联用技术测定饲料中己烯雌酚的方法,并对微萃取条件进行了优化。该方法在0.05～5.00mg/L范围内线性良好,相关系数(r)为0.998,检测限为0.5μg/kg,相对标准偏差小于10%。该方法绿色环保、简便、灵敏,可用于饲料中己烯雌酚的分析测定。     

Pharmacokinetics of ceftriaxone in Green sea turtles (Chelonia mydas) following intravenous and intramuscular administration at two dosages

Green sea turtles are widely distributed in tropical and subtropical waters. Adult green sea turtles face many threats, primarily from humans, including injuries from boat propellers, being caught in fishing nets, pollution, poaching, and infectious diseases. To the best of our knowledge, limited pharmacokinetic information to establish suitable therapeutic plans is available for green sea turtles. Therefore, the present study aimed to describe the pharmacokinetic characteristics of ceftriaxone (CEF) in green sea turtles, Chelonia mydas, following single intravenous and intramuscular administrations at two dosages of 10 and 25 mg/kg body weight (b.w.). Blood samples were collected at assigned times up to 96 hr. The plasma concentrations of CEF were measured by liquid chromatography tandem mass spectrometry. The concentrations of CEF in the plasma were quantified up to 24 and 48 hr after i.v. and i.m. administrations at dosages of 10 and 25 mg/kg b.w., respectively. The C_max values of CEF were 15.43 ± 3.71 μg/ml and 43.48 ± 4.29 μg/ml at dosages of 10 and 25 mg/kg, respectively. The AUC_last values increased in a dose‐dependent fashion. The half‐life values were 2.89 ± 0.41 hr and 5.96 ± 0.26 hr at dosages of 10 and 25 mg/kg b.w, respectively. The absolute i.m. bioavailability was 67% and 108%, and the binding percentage of CEF to plasma protein was ranged from 20% to 29% with an average of 24.6%. Based on the pharmacokinetic data, susceptibility break‐point and PK‐PD index (T > MIC, 0.2 μg/ml), i.m. administration of CEF at a dosage of 10 mg/kg b.w. might be appropriate for initiating treatment of susceptible bacterial infections in green sea turtles.     

A pharmacokinetic and residual study of sulfadiazine/trimethoprim in mandarin fish (Siniperca chuatsi) with single‐ and multiple‐dose oral administrations

A pharmacokinetic and tissue residue study of sulfadiazine combined with trimethoprim (SDZ/TMP = 5/1) was conducted in Siniperca chuatsi after single‐ (120 mg/kg) or multiple‐dose (an initial dose of 120 mg/kg followed by a 5‐day consecutive dose of 60 mg/kg) oral administrations at 28 °C. The absorption half‐life (t_1/2α), elimination half‐life (t_1/2β), volume of distribution (V_d/F), and the total body clearance (Cl_B/F) for SDZ and TMP were 4.3 ± 1.7 to 6.3 ± 1.8 h and 2.4 ± 1.0 to 3.9 ± 0.9 h, 25.9 ± 4.5 to 53.0 ± 5.6 h and 11.8 ± 3.5 to 17.1 ± 3.4 h, 2.34 ± 0.78 to 3.67 ± 0.99 L/kg and 0.39 ± 0.01 to 1.33 ± 0.57 L/kg, and 0.03 ± 0.01 to 0.06 ± 0.01 L/kg·h and 0.02 ± 0.01 to 0.05 ± 0.01 L/kg·h, respectively, after the single dose. The elimination half‐life (t_1/2β) and mean residue time (MRT) for SDZ and TMP were 68.8 ± 7.8 to 139.8 ± 12.3 h and 34.0 ± 5.5 to 56.1 ± 6.8 h, and 99.3 ± 6.1 to 201.7 ± 11.5 h and 49.1 ± 3.5 to 81.0 ± 5.1 h, respectively, after the multiple‐dose administration. The daily oral SDZ/TMP administration might cause a high tissue concentration and long t_1/2β, thereby affecting antibacterial activity. The withdrawal time for this oral SDZ/TMP formulation (according to the accepted guidelines in Europe for maximum residue limits, <0.1 mg/kg of tissues for sulfonamides, and <0.05 mg/kg for TMP) should not be <36 days for fish.     

Ketoprofen pharmacokinetics of R‐ and S‐isomers in juvenile loggerhead sea turtles (Caretta caretta) after single intravenous and single‐ and multidose intramuscular administration

Ketoprofen is a nonsteroidal anti‐inflammatory and analgesic agent that nonselectively inhibits cyclooxygenase, with both COX‐1 and COX‐2 inhibition. Recent studies on COX receptor expression in reptiles suggest that nonselective COX inhibitors may be more appropriate than more selective inhibitors in some reptiles, but few pharmacokinetic studies are available. The goal of this study was to determine single‐ and multidose (three consecutive days) pharmacokinetics of racemic ketoprofen administered intravenously and intramuscularly at 2 mg/kg in healthy juvenile loggerhead turtles (Caretta caretta). The S‐isomer is the predominant isomer in loggerhead sea turtles, similar to most mammals, despite administration of a 50:50 racemic mixture. Multidose ketoprofen administration demonstrated no bioaccumulation; therefore, once‐daily dosing will not require dose adjustment over time. S‐isomer pharmacokinetic parameters determined in this study were C_max of 10.1 μg/ml by IM injection, C₀ of 13.4 μg/ml by IV injection, AUC of 44.7 or 69.4 μg*hr/ml by IM or IV injection, respectively, and T½ of 2.8 or 3.6 hr by IM or IV injection, respectively. Total ketoprofen plasma concentrations were maintained for at least 12 hr above concentrations determined to be effective for rats and humans. A dose of 2 mg/kg either IM or IV every 24 hr is likely appropriate for loggerhead turtles.     

Association of single‐nucleotide polymorphism of melanocortin gene with feed intake in rabbit (Oryctolagus cuniculus)

The aim of the study was to investigate the association of two parts of melanocortin gene (MC4R‐1, MC4R‐2) and feed intake for V‐line rabbits. V‐line rabbits were grouped into high and low daily feed intake during the period from 30 to 63 days of age in order to identify MC4R SNPs useful for association study with feed intake. DNA from blood samples of each group was extracted to amplify the MC4R gene. The purified PCR products were sequenced in those had the highest and lowest feed intake. Alignment of sequence data from each group revealed that there is a variation detected in MC4R‐1 at nucleotide 35 (T‐G) (sense mutation) and another variation was detected in MC4R‐2 gene at nucleotide 19 (T‐C) (sense mutation) for high feed intake rabbits. These sense mutations lead to transform some amino acids and cause a significant change of the MC4R function. The results of average daily feed intake (ADFI) indicated that group (1) had significantly higher feed intake than group (2) of V‐line rabbits. The detected mutations and the analysis of daily feed intake means revealed a significant association between MC4R polymorphism and feed intake in rabbits.     

Conversion of α‐linolenic acid to long‐chain omega‐3 fatty acid derivatives and alterations of HDL density subfractions and plasma lipids with dietary polyunsaturated fatty acids in Monk parrots (Myiopsitta monachus)

The effect of α‐linolenic acid from a flaxseed (FLX)‐enriched diet on plasma lipid and fatty acid metabolism and possible atherosclerosis risk factors was studied in Monk parrots (Myiopsitta monachus). Twenty‐four Monk parrots were randomly assigned to diets containing either 10% ground SUNs or 10% ground FLXs. Feed intake was calculated daily. Blood samples, body condition scores and body weights were obtained at ?5 weeks, day 0, 7, 14, 28, 42 and 70. Plasma samples were analysed for total cholesterol, free cholesterol, triacylglycerols and lipoproteins. Phospholipid subfraction fatty acid profiles were determined. By day 70, the FLX group had significantly higher plasma phospholipid fatty acids including 18:3n‐3 (α‐linolenic acid), 20:5n‐3 (eicosapentaenoic acid) and 22:6n‐3 (docosahexaenoic acid). The sunflower group had significantly higher plasma phospholipid levels of 20:4n‐6 (arachidonic acid). By day 70, the high‐density lipoprotein (HDL) peak shifted resulting in significantly different HDL peak densities between the two experimental groups (1.097 g/ml FLX group and 1.095 g/ml SUN group, p = 0.028). The plasma fatty acid results indicate that Monk parrots can readily convert α‐linolenic acid to the long‐chain omega‐3 derivatives including docosahexaenoic acid and reduce 20:4n‐6 accumulation in plasma phospholipids. The reason for a shift in the HDL peak density is unknown at this time.