Pharmacokinetics of Florfenicol after Oral Administration in Yellow Catfish,Pelteobagrus fulvidraco

The pharmacokinetics of florfenicol (FF) and its metabolite, florfenicol amine (FFA), were investigated after doses of 10 mg/kg/day were administered orally per os (p.o.) on a single day or on three consecutive days in yellow catfish, Pelteobagrus fulvidraco, raised in water temperatures of 25 C. After a single dose p.o. was administered, the apparent volume of distribution at steady state (V_dss) of FF was computed to be 2.52 L/kg. The T_max values were in the following order: liver (1.82 h) < kidney (2.26 h) < skin (6.15 h) < muscle (6.32 h) < plasma (7.25 h). These results show that FF and FFA accumulated rapidly in the kidney and liver. The t_1/2β values in plasma, muscle, skin, liver, and kidney were 9.63, 15.75, 14.44, 11.55, and 15.75 h, respectively, for FF and 21.66, 15.07, 17.33, 26.65, and 30.13 h, respectively, for FFA. After a single p.o. dose was administered on three consecutive days, the t_1/2β values of FF and FFA in skin‐on muscle were 17.12 and 13.55 h, respectively. The total concentrations of FF and FFA in skin‐on muscle 1, 3, and 5 d after the last administration were 3.39, 0.5, and 0.062 µg/g, respectively.     

Pharmacokinetic–pharmacodynamic modelling for the determination of optimal dosing regimen of florfenicol in Nile tilapia (Oreochromis niloticus) at different water temperatures and antimicrobial susceptibility levels

Optimized dosing regimen is key to the effective use of antibacterials and to minimizing drug‐related side effects. The current study established a pharmacokinetic–pharmacodynamic (PK‐PD) model for the determination of optimal antibacterial dosing regimen in fish taken into consideration the temperature‐dependent PK and the pathogen‐dependent antimicrobial susceptibility, using florfenicol (FF) in Nile tilapia as an example. The calculated optimal dosages significantly varied by temperature and target MIC levels, ranging from 2.23 (MIC 1 µg/ml at 24°C) to 34.88 mg kg^?1 day^?1 (MIC 4 µg/ml at 32°C). The appropriateness of the calculated dosages was successfully verified by the in vivo studies. After 5 days of oral administration of the calculated optimal dosage at 24°C, the predicted plasma drug values were in line with the mean observed C_min(ss) while at 28 and 32°C underestimation of the C_min(ss) in a dose‐dependent manner was observed and likely due to the occurrence of non‐linear PK at high dosages. The averaged serum protein binding of FF was 19.1%. Our results demonstrated the appropriateness and clinical applicability of the developed PK‐PD approach for the determination of optimal dosing regimens at given temperatures and MICs. Saturation metabolism and PK non‐linearity of FF in tilapia warrant further study.     

Accumulation of heavy metals and total phosphorus in intensive aquatic farm sediments: comparison of tilapia Oreochromis niloticus×Oreochromis aureu,Asian seabass Lates calcarifer and white shrimp Litopenaeus vannamei farms

This paper compares and discusses the accumulation of eight heavy metals (Cd, Co, Cr, Cu, Pb, Fe, Mn and Zn) and total phosphorus (TP) in surface sediments from five intensive farms of tilapia Oreochromis niloticus×Oreochromis aureu, Asian seabass Lates calcarifer and white shrimp Litopenaeus vannamei (n=75). Results indicate that the sediments were significantly contaminated by Cd, Co, Cr, Cu, Pb, Fe, Mn and Zn as well as P. The mean concentrations of Cd (0.47 mg kg^?1), Co (2.25 mg kg^?1), Cu (31.56 mg kg^?1), Pb (23.45 mg kg^?1), Fe (1.15%), Mn (145.5 mg kg^?1), Zn (41.5 mg kg^?1) and TP (750.69 mg kg^?1) in tilapia farm surface sediments were significantly lower than the levels observed in Asian seabass and white shrimp farm surface sediments, where the concentrations were, respectively, Cd – 2.69 and 2.35 mg kg^?1, Co – 17.66 and 15.22 mg kg^?1, Cu – 116.84 and 110.03 mg kg^?1, Pb – 38.5 and 33.26 mg kg^?1, Fe – 3.4% and 3.31%, Mn – 853.61 and 728.5 mg kg^?1, Zn – 105.21 and 83.92 mg kg^?1 and TP – 1445.63 and 1069.8 mg kg^?1. The heavy metal content of the sediments follow the order: Asian sea bass>white shrimp>tilapia. Cadmium and Cu in Asian seabass and white shrimp farm sediments would pose a major potential risk. Further, the sediment is highly enriched by Fe, P, Mn and Co.     

Pharmacokinetic/pharmacodynamic properties and P‐glycoprotein expression in pacific white shrimp,Litopenaeus vannamei after oral administration at three dosages of oxolinic acid

The experiments explored the pharmacokinetics (PK) properties of oxolinic acid (OXA) after oral administration at three dosages (10, 30 and 80 mg/kg) via medicated feed in the shrimp. The results showed that the C_max values of 4.31, 14.93 and 16.62 mg/L and AUC_0–∞ values of 92.61, 252.30 and 364.27 mg hr^?1 L^?1 were observed at three OXA dosage groups in the haemolymph respectively. In the hepatopancreas, C_max values of 7.90, 27.23 and 60.51 mg/kg and AUC_0–∞ values of 42.01, 133.06 and 219.06 mg hr^?1 L^?1 were observed at 0.5 hr post administration respectively. In the muscle, C_max values of 1.62, 5.80 and 7.36 mg/kg and the AUC_0–∞ values of 25.64, 98.10 and 134.24 mg hr^?1 L^?1 were observed at 2 hr post administration respectively. In the gills, C_max values of 2.87, 8.08 and 12.12 mg/kg and the AUC_0–∞ values of 51.38, 118.65 and 206.48 mg hr^?1 L^?1 were observed at 4 hr post administration respectively. In addition, the in vitro MIC values of OXA at three dosages against 132 strains of Vibrio were examined and showed that the minimum inhibitory concentration (MIC) values for OXA primarily ranged from 0.15–1.25 µg/ml, including eight strains of Vibrio showing MIC values ≥5 µg/ml. The MIC₅₀ and MIC₉₀ values of 132 strains were 0.62 and 1.25 μg/ml respectively. The AUC_0–24/MIC₉₀ ratios of Vibrio were 140.4 in 30 mg/kg group. Furthermore, the P‐glycoprotein (P‐gp) expression was determined in shrimp tissues after administration to three dosage groups (10, 30 and 80 mg/kg). The results showed that P‐gp expression was up‐regulated in the hepatopancreas (5.36‐, 13.68‐ and 31.06‐fold respectively) compared with the control group.     

高效液相色谱法检测克氏原螯虾组织中氟苯尼考及氟苯尼考胺

建立了克氏原螯虾(Procambarus clarkii)血淋巴、肌肉、肝胰脏中氟苯尼考及氟苯尼考胺的高效液相色谱检测方法。采用Agilent Eclipse XDB-C18(250mm×4.6 mmi.d,5μm)反相色谱柱,以A液(乙腈):B液(0.04 mol/L NaH2PO4、0.004 mol/L十二烷基硫酸钠和0.10%三乙胺)(体积比为30∶70)为流动相,225 nm紫外检测波长完成分析。结果显示:本方法在0.025~20 mg/L范围内浓度与峰面积呈良好的线性关系;组织的加标回收率为78.8%~94.2%,相对标准偏差小于8.16%,氟苯尼考在血淋巴、肌肉、肝胰脏的检出限分别为0.01 mg/L、0.02 mg/kg、0.03 mg/kg,氟苯尼考胺则为0.015 mg/L、0.03 mg/kg、0.05 mg/kg。     

Determination of Dose Rate of Florfenicol in Feed for Control of Mortality in Channel Catfish Ictalurus punctatus (Rafinesque) Infected with Edwardsiella ictaluri, Etiological Agent of Enteric Septicemia

–A dose titration study was conducted to determine the appropriate dosage of florfenicol in feed to control mortality in channel catfish Ictalurus punctatus associated with enteric septicemia of catfish caused by Edwardsiella ictaluri. Six tanks (20 fish/ tank) were assigned to each of the following treatment: 1) not challenged with E. ictaluri and fed unmedicated feed; 2) challenged with E. ictaluri and fed unmedicated feed; 3) challenged and fed 5-mg florfenicol/kg body weight (kg bw); 4) challenged and fed 10-mg florfenicol/kg bw; or 5) challenged and fed 15-mg florfenicol/kg bw. Treatment was initiated the day after inoculation, and feed was administered by hand at 2.5% body weight for 10 consecutive days. Feeding activity was scored for all groups and was noted to be significantly less than the challenged, unmedicated group. Cumulative mortality in the challenged untreated group was 60%. The mortality in the unchallenged untreated group was 0%, and in die 5-, 10-, 15-mg florfenicol/kg bw group was 2.5%, 0.8%, and 2.5%, respectively. The mortality in each challenged, treated group and the non-challenged control group was significantly less than the challenged, unmedicated controls (P < 0.0001 for each contrast). There were no pairwise statistically significant contrasts among the florfenicol treated groups and the non-challenged control group. All 600 fish in the study were necropsied, cultured for bacteria, and examined by gross pathology. No specific lesions that could be associated with the antibiotic were observed. The efficacy of the 10 mg/kg dosage was confirmed in a separate dose confirmation study. In this study, fish in 30 tanks (20 fish/ tank) were infected with E. icraluri by immersion. Two days post-inoculation, fish in 15 tanks were hand-fed unmedicated feed, and 15 tanks were hand-fed medicated feed at a dosage of 10-mg florfenicol/kg bw at 2.5% body weight for 10 d. Feeding activity was scored and was noted to be significantly less than the challenged, unmedicated group. Cumulative mortality in the florfenicol group (14%) was significantly less than cumulative mortality in the untreated group (87.3%) (P < 0.0001). All 600 fish were submitted for bacterial culture, necropsied. and examined for gross pathology, and once again, no specific lesions that could be associated with the antibiotic were observed. The minimum inhibitory concentration of florfenicol against E. ictaluri in both studies was 0.25 ug/mL. Florfenicol was palatable, safe, and efficacious for control of mortality due to infection by E. ictaluri in catfish.     

Vibrio alginolyticus infection in Asian seabass (Lates calcarifer,Bloch) reared in open sea floating cages in India

Asian seabass (Lates calcarifer) has been designated as a candidate species for open sea cage culture in India. A case of vibriosis in Asian seabass reared in open sea floating cages is reported. Haemorrhage and ulcer were observed grossly in the diseased fish. The pathogen, identified as Vibrio alginolyticus based on biochemical and molecular characterization, was isolated from liver, gill, kidney, brain and blood. Histological examination of the diseased fish showed congestion, haemorrhage and necrosis in vital organs. The LD₅₀ studies showed that the organism was virulent to Asian seabass, and the LD₅₀ value was 10^3.2 CFU g^?1 fish.     

盐酸土霉素在奥尼罗非鱼体内的药代动力学及残留研究

在水温(25±2)℃条件下,以15 mg/kg鱼体重的剂量给奥尼罗非鱼单次口灌盐酸土霉素,采用高效液相色谱法测定血浆和肌肉组织中的药物浓度,研究盐酸土霉素在奥尼罗非鱼体内的代谢及消除规律。结果显示:血药时间数据符合一级吸收二室开放模型,半衰期(T1/2Ka、T1/2α、T1/2β)分别为4.79、4.10、45.20 h,最大血药浓度为1.50μg/m L,达峰时间为7.30 h,药时曲线下面积(AUC)为42.35μg·h/m L。肌肉作为可食性组织,选取肌肉组织作为残留检测的靶组织,以0.1 mg/kg为最高残留限量,在本试验条件下,建议休药期不低于10 d。     

Assessment of Aquaflor(®) , copper sulphate and potassium permanganate for control of Aeromonas hydrophila and Flavobacterium columnare infection in sunshine bass, Morone chrysops female × Morone saxatilis male

Two experiments were conducted to test the effectiveness of different therapeutants against a mixed infection of Aeromonas hydrophila and Flavobacterium columnare in sunshine bass. Experiment 1 evaluated copper sulphate, florfenicol‐medicated feed and potassium permanganate (KMnO₄) against a natural mixed infection. Experiment 2 further evaluated copper sulphate as a treatment to control an experimental mixed infection. In experiment 1, naturally infected untreated fish had the lowest final survival per cent, at 71%, while florfenicol‐medicated feed at 15 mg kg^?1 body weight for 10 days or copper sulphate at 2.1 mg L^?1 (1% of the total alkalinity) for 24 h produced the highest final survivals, at 90% and 88%, respectively. The final survival of the naturally infected fish administered florfenicol‐medicated feed was significantly different (P < 0.1) from the untreated fish. The survival curves for the florfenicol and the copper sulphate at 2.1 mg L^?1 were significantly improved from the untreated fish. In experiment 2, fish were challenged by waterborne exposure to A. hydrophila and F. columnare and either not treated or treated with copper sulphate at 2.1 mg L^?1. At the end of experiment 2, the per cent survival of the challenged fish treated with copper sulphate (99%) was significantly higher (P < 0.05) than the non‐treated (61%). The results illustrate clear benefit of florfenicol and copper sulphate against a mixed infection of A. hydrophila and F. columnare.     

Dietary administration of astaxanthin improves feed utilization,growth performance and survival of Asian seabass,Lates calcarifer (Bloch, 1790)

This study was undertaken to assess the impacts of dietary astaxanthin supplementation on growth performance, feed utilization, survival, and serum growth hormone (GH) availability of Asian seabass, Lates calcarifer, with special reference to dose–response relationships and variations during different feeding phases (short‐term, medium‐term and long‐term). Fish were fed the following diets in triplicate for 90 days: the control (CD), AX50 (50 mg astaxanthin/kg diet), AX100 (100 mg astaxanthin/kg diet) and AX150 (150 mg astaxanthin/kg diet). The findings revealed that fish exhibited significant linear increments (p < .05) in specific growth rate (SGR), weight gain, feed utilization efficiency and survival when fed various diets with escalating levels of astaxanthin. Supplementation with dietary astaxanthin significantly augmented (p < .05) GH levels in fish. Significant positive associations (p < .05) were observed between circulating serum GH levels and SGR of fish from all groups following three consecutive feeding phases, denoting a robust cause‐and‐effect relationship. Circulating GH concentrations were considered as a sensitive biomarker of growth performance in Asian seabass. This study illustrated that supplemental astaxanthin could be administered in culture protocols to improve the growth rate and commercial hatchery production of Asian seabass, and possibly other teleost species.     

Effects of florfenicol feeding on diversity and composition of the intestinal microbiota of channel catfish (Ictalurus punctatus)

Intestinal microbiota contributes health of living organisms. Florfenicol is an approved antimicrobial (AM) prescribed for several bacterial fish diseases. The present study investigated the extent to which florfenicol modulates intestinal microbial populations of channel catfish (Ictalurus punctatus). Florfenicol was administered orally to catfish at a standard therapeutic dose (10–15 mg/kg of body weight for 10 days), and the intestinal contents were collected and the 16S rRNA was subjected to Illumina sequencing. Alpha diversity analysis indicated that florfenicol significantly decreased microbiota richness and diversity. Beta diversity reflected a clear separation had occurred between the florfenicol‐fed and control groups. Results indicated a significant increase in the abundance of phylum Proteobacteria (98.97% vs. 79.35% of the population) and decrease in phyla Firmicutes and Bacteroidetes (0.72% and 0.29% vs. 18.04% and 2.53%, respectively) in the florfenicol‐fed fish in comparison with the control fish. At the genus level, unclassified Enterobacteriaceae and Escherichia populations increased in the florfenicol group. In contrast, Plesiomonas, Aeromonas, Lactococcus, Clostridium sensu stricto, Romboutsia, Klebsiella, Turicibacter and Lactobacillus decreased in florfenicol‐fed fish in comparison with the control fish, indicating that intestinal microbiota of catfish was substantially modulated by florfenicol administration. Knowledge of changes in gut microbiota during medicated feed administration is important to improve fish performance and disease management and could enable the development of alternative therapeutic strategies.     

Determination of the muscle residue levels and withdrawal time of florfenicol in healthy and experimentally Lactococcus garvieae infected rainbow trout (Oncorhynchus mykiss,Walbaum 1792) grown in the Black Sea water

The aim of this study was to determine, muscle residue level (MRL) and withdrawal time (WT) of florfenicol (FF), in healthy and infected rainbow trout (Oncorhynchus mykiss) with Lactococcus garvieae. Fish were divided into the four groups, Group I (Infected fish fed with FF free diet, n = 10), Group II (Healthy fish fed with FF free diet, n = 10), Group III (Infected fish fed with FF containing diet, n = 80) and Group IV (Healthy fish fed with FF containing diet, n = 80). Fish in Group I and Group III were infected with 1.8×10⁶ cfu/ml Lactococcus garvieae by immersion. FF containing diet was applied fish (10 mg kg^‐1 day^‐1) in Group III and IV for 10 days. FF residues in fish muscles were analysed by validated HPLC method. The limit of detection (LOD) and limit of quantification (LOQ) values of the method were determined to be 0.31 μg/g and 0.64 μg/g, respectively, and the recovery was 92.61%. As a result of the experiments, there was no significant difference between the infected and healthy group in term of MRLs. However, in both groups, it was determined that third‐day values were below MRLs (1,000 μg/kg). Also, the WT values were lower than the 500°C‐day values reported in the EC commission decision in both groups and were determined as 14.5 and 14.12 (water temperature mean 18°C ± 1) per day, respectively.     

Pharmacokinetic model of florfenicol in turbot (Scophthalmus maximus): establishment of optimal dosage and administration in medicated feed

The pharmacokinetics of florfenicol (FF) in turbot (Scophthalmus maximus) was studied after single intravenous (10 mg kg⁻¹) and oral (100 mg kg⁻¹) administration. The plasma concentration–time data of florfenicol were described by an open one‐compartment model. The elimination half‐life (t_1/2) was estimated to be 21.0 h, and the total body clearance, Cl, was determined as 0.028 L kg h⁻¹. The apparent volume distribution (V_d) was calculated to be 0.86 L kg⁻¹ and the mean residence time (MRT_iv) was 30.2 h. Following oral administration, the maximum plasma concentration (C_max) of 55.4 μg mL⁻¹ was reached at 12 h (T_max). The absorption constant (k_a) was 0.158 h⁻¹. The bioavailability was estimated to be 57.1%. The low bioavailability observed at higher doses was explained by the saturation of the mechanisms of absorption. The drug absorption process was limited by its inherent low solubility, which limited the amount of available FF absorbed in the gastrointestinal tract. Based on the pharmacokinetic data, an optimal dosing schedule for FF administration is hereby provided. Based on the minimum inhibitory concentration found for susceptible strains of Aeromonas salmonicida, oral FF administration of first, an initial dose of 30 mg FF kg⁻¹, followed by 6 maintenance doses at 18 mg kg⁻¹/daily could be effective against furunculosis in turbot.     

Comparison of the pathogenicity of Francisella orientalis in Nile tilapia (Oreochromis niloticus), Asian seabass (Lates calcarifer) and largemouth bass (Micropterus salmoides) through experimental intraperitoneal infection

Francisella orientalis is a highly virulent, emerging bacterium that causes mass mortalities in tilapia. This pathogen also affects numerous other warm-water fish species, including three-line grunt, hybrid striped bass and various ornamental fish. This study sheds light on two new species of fish that are susceptible to F. orientalis. Asian seabass and largemouth bass showed variable levels of susceptibility in a bacterial challenge experiment. After intraperitoneally injected with a dose of 10⁶ CFU/fish, a total of 64.28% and 21.42% mortalities were obtained in Asian seabass and largemouth bass, respectively. Meanwhile, Nile tilapia showed acute mortality of 100%. All fish showed typical lesions of francisellosis, including multifocal granulomas in the spleen and head kidney. Immunohistochemical analysis revealed strong positive signals inside the granulomas of all fish. The bacterial recovery in solid media from infected fish was highest in Nile tilapia (85.71%), followed by Asian seabass (35.71%) and largemouth bass (21.42%). PCR results tested 100% positive for Nile tilapia, and 78.57% and 21.42% for Asian seabass and largemouth bass, respectively. In conclusion, Asian seabass and largemouth bass are susceptible to this pathogen, which warrants new management strategies when employing predation polyculture systems of these species with tilapia.     

Depletion of florfenicol amine in tilapia (Oreochromis sp.) maintained in a recirculating aquaculture system following Aquaflor®‐medicated feed therapy

Aquaflor^® [50% w w^?1 florfenicol (FFC)], is approved for use in freshwater‐reared warmwater finfish which include tilapia Oreochromis spp. in the United States to control mortality from Streptococcus iniae. The depletion of florfenicol amine (FFA), the marker residue of FFC, was evaluated after feeding FFC‐medicated feed to deliver a nominal 20 mg FFC kg^?1 BW d^?1 dose (1.33× the label use of 15 mg FFC kg^?1 BW d^?1) to Nile tilapia O. niloticus and hybrid tilapia O. niloticus × O. aureus held in a recirculating aquaculture system (RAS) at production‐scale holding densities. Florfenicol amine concentrations were determined in fillets taken from 10 fish before dosing and from 20 fish at nine time points after dosing (from 1 to 240 h post‐dosing). Water samples were assayed for FFC before, during and after the dosing period. Parameters monitored included daily feed consumption and biofilter function (levels of ammonia, nitrite and nitrate). Mean fillet FFA concentration decreased from 13.77 μg g^?1 at 1‐h post dosing to 0.39 μg g^?1 at 240‐h post dosing. Water FFC concentration decreased from a maximum of 1400 ng mL^?1 at 1 day post‐dosing to 847 ng mL^?1 at 240 h post‐dosing. There were no adverse effects noted on fish, feed consumption or biofilter function associated with FFC‐medicated feed administration to tilapia.     

Depletion study and estimation of withdrawal periods for florfenicol and florfenicol amine in pacu (Piaractus mesopotamicus)

The intensive production of farmed fish is at a global all‐time high, and the control of bacteria proliferation in fish farms requires the frequent use of antimicrobials. This practice raises important environmental concerns related to the emergence of antimicrobial‐resistant bacteria strains. Only a few antimicrobial drugs have been approved for use in aquaculture, one of which is florfenicol. This work studies the depletion and withdrawal period of florfenicol and its main metabolite, florfenicol amine, in pacu (Piaractus mesopotamicus), a neotropical characin widely farmed in the southern hemisphere. Juvenile pacu (average weight of 724 g) were stocked in a closed‐loop laboratory system with controlled water temperature (25.8°C), and were fed for 10 consecutive days with a diet containing an intended dose of 10 mg/florfenicol per kg bw. Muscle and skin tissues were collected at 1, 3, 6, 8, 10, 12 and 16 days post‐treatment, and florfenicol and florfenicol amine were quantified using a validated ultra‐high performance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) method. The limits of quantitation for florfenicol and florfenicol amine were 10 ng/g in muscle and 50 ng/g in skin. Considering a maximum residue limit of 1000 ng/g for the sum of florfenicol and florfenicol amine in muscle with skin in natural proportions a withdrawal period of 5 days (water temperature 25.8°C) or 129 degree days was calculated on the basis of the upper limit of the one‐sided 95% confidence interval for the 99^th percentile derived from the residue depletion study.     

Erythromycin and florfenicol treatment of rainbow trout Oncorhynchus mykiss (Walbaum) experimentally infected with Flavobacterium psychrophilum

Flavobacterium psychrophilum is responsible for significant economic losses in rainbow trout aquaculture. Antimicrobial treatment remains the primary means of control; however, there are limited choices available for use. The objectives of the study were therefore to determine the minimum inhibitory concentrations for erythromycin and florfenicol in selected F. psychrophilum isolates and to evaluate their clinical treatment efficacy in experimentally infected rainbow trout. All isolates tested had moderate susceptibility to florfenicol and erythromycin except one isolate, which had low susceptibility to erythromycin. Two isolates (one with moderate and one with low susceptibility to erythromycin) were used in an experimental infection trial. Rainbow trout juveniles were injected intraperitoneally with 10⁸ cfu/fish and after mortality had begun, fish were given erythromycin‐ and florfenicol‐medicated feed at a rate of 75 mg kg^?1 day^?1 and 10 mg kg^?1 day^?1 fish body weight, respectively, for 10 consecutive days. The splenic F. psychrophilum load was determined using an rpoC quantitative PCR throughout the 30‐day trial. Relative to antibiotic‐free controls, erythromycin treatment significantly (p < 0.05) reduced mortality of rainbow trout juveniles infected with FPG101, even when treatment was initiated after clinical signs developed.     

模拟探究凡纳滨对虾磷脂在贮藏过程中的水解机理

为探究凡纳滨对虾磷脂(PL)在贮藏过程中的水解机理,本研究建立了磷脂相关酶与磷脂酰胆碱(PC)的体外模型,模拟凡纳滨对虾磷脂水解反应。实验分别从凡纳滨对虾体内分离纯化得到磷脂相关酶与PC,进行体外反应,并采用"鸟枪"脂质组学法及气相色谱—质谱法分析水解反应前后PC、溶血磷脂(LPL)及游离脂肪酸(FFA)的组成和含量变化,进而推断磷脂的水解机理。结果显示,凡纳滨对虾体内磷脂水解酶包括脂肪酶、磷脂酶A_1(PLA_1)、磷脂酶A_2(PLA_2)、磷脂酶C(PLC)及磷脂酶D(PLD),其中PLA2活力最高,可达177.87 U。体外模拟反应中,凡纳滨对虾PC含量由516.45显著降至146.14 mg/g,总FFA含量由36.42上升至568.57 mg/g,其中多不饱和脂肪酸(PUFA)显著增加了280.5 mg/g。通过相关性分析水解前后磷脂水解酶活力变化与PC含量变化,发现PC的变化与PLA2显著相关(R=0.91)。研究表明水产品中PC变化是磷脂相关酶催化的水解反应,其水解产物主要为FFA与LPC,且PLA_2对水产品贮藏过程中磷脂的水解影响最大。本实验建立了新型的体外水解模型,并初步探究了磷脂的水解机理,为水产品贮藏及磷脂酶的研究及应用奠定了理论基础。     

Susceptibility of freshwater rearing Asian seabass (Lates calcarifer) to pathogenic Streptococcus iniae

Asian seabass (Lates calcarifer) has been recognized as an economically important aquaculture species which can be adapted to and cultivated in wide range of salinities. The number of freshwater intensive seabass farms in Thailand is increasing annually. Here, we first describe the susceptibility of Asian seabass, which were cultured in freshwater, to Streptococcus inae (SI) and their pathological changes. Three isolates of putative SI were identified using a combination of standard biochemical assays and species‐specific PCR prior subjected to in vivo challenge. Accumulated mortalities of the fish which received 10⁷ CFU fish^?1 of either SI1J, SI SGSA or SI2J were 90%, 90% and 100% at 7 days‐post infection (dpi), respectively, and mortalities increased sharply between 3 and 5 dpi. Clinical signs such as erratic swimming and opaque eyes were identified from a few infected fish, while most died rapidly without any abnormal signs. Histopathological manifestations were observed in the multiple organs (kidney, liver and brain). Haemorrhage, hyperhemia, cellular degeneration and inflammatory cells infiltration were commonly found within the internal organs. Notably, the formation of numerous encyst‐like lesion aggregated by eosinophilic cells, resembling macrophages, were typically found in the brain of the infected fish. Summarily, this study first revealed that freshwater reared Asian seabass is highly susceptible to SI infection and haemorrhagic septicaemia was a major pathological change that could be found in the infected fish.