Pharmacokinetics of oxolinic acid and oxytetracycline in kuruma shrimp, Penaeus japonicus

The pharmacokinetics of oxolinic acid and oxytetracycline were examined in kuruma shrimp (Penaeus japonicus) after intra-sinus (10 and 25 mg/kg, respectively) and oral (50 mg/kg) administration. The shrimp were kept in tanks with recirculated artificial seawater at a salinity of 22–23 ppt. The water temperature was maintained at 25±0.6 °C. The hemolymph concentrations of both drugs after intra-sinus dosing were best described by a two-compartment open model. The distribution and elimination half-lives (t_1/2 and t_1/2β) were found to be 0.59 and 33.2 h for oxolinic acid and 0.45 and 24.7 h for oxytetracycline, respectively. The apparent volume of distribution at a steady state (V_ss) and total body clearance (CL_b) were estimated to be 1309 ml/kg and 28.8 ml/kg/h for oxolinic acid and 748 ml/kg and 22.7 ml/kg/h, respectively. The hemolymph concentration–time curves after oral administration did not fit by the nonlinear least squares method using one- and two-compartment model with first-order absorption in either of the drugs. The peak hemolymph concentration (C_max), the time to peak hemolymph concentration (t_max) and the elimination half-life were found to be 17.8 μg/ml, 7 h and 34.3 h for oxolinic acid and 24.3 μg/ml, 10 h and 33.6 h for oxytetracycline, respectively. The bioavailability (F) after oral administration was 32.9% for oxolinic acid and 43.2% for oxytetracycline. The hemolymph protein binding in vivo was determined to be 36.7±8.5% for oxolinic acid and 22.9±4.8% for oxytetracycline.     

The effectiveness of a commercial microbial product in poorly prepared tiger shrimp, Penaeus monodon (Fabricius), ponds

The efficacy of a commercial microbial product was tested in commercial tiger shrimp, Penaeus monodon (Fabricius), ponds for one culture period in Kuala Selangor, Malaysia. Four ponds with replicates for treatment and control were used. The pond bottom was dried but the organic sludge was not removed as normally practised in pond preparation. The ponds were stocked with 15 post‐larvae at the rate of 31.m^?2. Physical, chemical and biological parameters of the pond were analysed every 2 weeks during the culture period. Water quality parameters remained within the optimum range for shrimp culture except for ammonia‐nitrogen being significantly higher in control ponds and silica in treated ponds. Benthic organisms were not found in any of the ponds. The average counts of different bacteria were not significantly higher in treated ponds than control. Because of poor health, the shrimp were harvested earlier (72 days) than the usual 120 days. An average of 875.60 ± 67.00 kg shrimp ha^?1 was obtained in treated ponds with a feed conversion ratio (FCR) of 1.57 ± 0.10 and survival rate of 42.35 ± 5.37% compared with 719.50 ± 130.94 kg shrimp ha^?1, 2.99 ± 0.70 and 21.25 ± 3.26%, respectively, in control ponds. Neither the microbial product nor the frequent water exchange was effective in overcoming the problems caused by the poor pond bottom.     

The study on the cytology and histochemistry of lymphoid organ spheroids in Penaeus chinensis

The lymphoid organ spheroids (LOS) were investigated by cytological and histochemical techniques in Penaeus chinensis. The results showed that the LOS only appeared in the individuals with relatively large lymphoid organ (long diameter>2 mm.). The LOS were mainly made up of two types of cells, one of which had a low cytoplasm to nuclear volumetric ratio, developed marginated chromatin and 1–2 nucleoli, while the other had large cytoplasm to nuclear volumetric ratio, homogeneous chromatin, and no or only one nucleolus. These two kinds of cells all contained mitochondria and rough endoplasmic reticulum. The necrotic cells could usually be observed. The LOS showed negative PAS reaction, weak positive phenoloxidase, and negative -naphthyl-acetate esterase (-NAE).     

The effect of vitamin A supplementation in broodstock feed on reproductive performance and larval quality in Penaeus chinensis

The effect of feeding four semi‐purified diets A1, A2, A3, A4, containing different vitamin A acetate levels 0, 20, 40, 60 mg kg^?1 diet, respectively, on fecundity, egg hatching rate, larval survival rate and vitamin A content in eggs of Chinese shrimp (Penaeus chinensis) broodstock was compared with a fresh clam diet (control) in a 60‐day feeding trial. The broodstock shrimp fed the diet with 60 mg kg^?1 vitamin A acetate added exhibited significantly higher fecundity (P < 0.01). Hatching rate was highest with diet A4 (P < 0.05), whereas hatching rates were similar fed diets A1, A2, A3. Increasing levels of vitamin A in broodstock diet resulted in improvement in larval quality. The vitamin A levels in shrimp eggs from broodstock fed with diet A4 were higher compared with those from broodstock fed with diet A1, A2 (P < 0.01). The fecundity and hatching percentages were positively correlated with the vitamin A content in eggs in the present study. The results of this study showed that higher level of vitamin A in broodstock diet may have positive effects on fecundity and larval quality in P. chinensis.     

Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Characterization and application of monoclonal antibodies against white spot syndrome virus

Three hybridoma clones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with white spot syndrome virus (WSSV) isolated and purified from Penaeus monodon (Fabricius), collected from north-eastern Taiwan. By sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE), the protein profile of this isolate contained four major proteins with sizes of approximately 35 (VP35), 28 (VP28), 24 (VP24), and 19 kDa (VP19). Western blot analysis revealed that two MAbs (1D7 and 6E1) recognized epitopes on VP28 and one MAb (3E8) recognized an epitope on VP19. The MAb 6E1 isotyped to the IgG₁ class was used in both an indirect immunofluorescence assay (IFA) and in an immunochemical staining protocol for successful identification and localization of WSSV in infected shrimp tissues. Antigenic similarity of isolates from Indonesia and Malaysia to the Taiwan isolate was illustrated by IFA with MAb 6E1. A MAb (2F6) which bound specifically to two shrimp proteins, 75 and 72 kDa, and reacted to the healthy and non-target tissues of WSSV in infected shrimp, such as hepatopancreas, is also described here and shows the necessity for specific identification of antibodies.     

Histological examination of final oocyte maturation and atresia in wild and domesticated Penaeus monodon (Fabricius) broodstock

Oocyte maturation and gonadosomatic index (GSI) of eyestalk ablated Penaeus monodon females collected from the wild and from two first‐generation domesticated lines were assessed. Frequency and diameter of the different oocytes, and the intensity of oocyte atresia, were compared among groups through histological assessments of the sections of the middle ovarian lobe. Digitized images from ovary sections were used to record the frequency and diameter of different oocyte types. Spawning performance of the three groups were expressed in terms of the percentage of females that spawned at least once (productive females), time from eyestalk ablation to first spawning (latency period) and the number of spawnings per female stocked. Final ovarian maturation was attained in all groups, as indicated by the presence of mature oocytes with cortical rods (cortical oocytes), dark‐green ovarian colour and high GSI values (5.83–6.86%). However, domesticated groups presented significant larger immature oocyte types (previtellogenic and yolky oocytes) and smaller cortical oocytes compared with wild females, indicating a reduced vitellogenic activity during final oocyte maturation. Additionally, the frequency of atresia was comparatively higher for both domesticated groups, which could be related to their inferior spawning performance. The implications of these results on the reproductive potential and development of domesticated P. monodon are discussed.     

Use of ongrownArtemia in nursery culturing of the tiger shrimp

Juvenile and adultArtemia produced in a semi flow-through culture system were used as food for postlarval shrimp. The growth performance of shrimp reared on such ongrownArtemia live prey is identical to the growth obtained when feeding newly hatchedArtemia. However, a significantly better stress resistance is obtained when the postlarvae are exposed to a low salinity in a stress test. Besides nutritional and energetic advantages, the use ofArtemia biomass for feeding postlarval shrimp also results in improved economics as expenses for cysts and weaning diets can be reduced.     

斑节对虾基因组微卫星分离及其序列特征研究

采用(CA)_(12)(AG)_(12)及(TA)_(16)生物素标记探针及磁珠富集法构建了斑节对虾Penaeus monodon基因组微卫星富集文库。随机挑选254个克隆进行PCR筛选,得到51个候选克隆(20．1%)。其中,32个克隆来源于CA-文库,另19个克隆来源于AG-文库。测序发现48个克隆含有微卫星重复单元,通过序列比对,最终获得40个具有特异微卫星序列的阳性克隆。微卫星(GA/CT)_N及(CA/GT)_n 2碱基重复序列分别占所有分离的微卫星数目的20．7%及60．4%。此外,还检测到其它多种微卫星重复类型,如(AT)_n、(GC)_n、(TGG)_n、(AAG)_n、(AAT)_n、(GAA)_n、(GTGC)_n、(GCGT)_n、(GGTTA)_n、(GTGCGT)_n,占检测到的微卫星数目的18．9%。获得的微卫星序列中属于完全型序列的有76条(68．5%),不完全型序列的有22条(19．8%),另有13条属于复合型序列(11．7%)。微卫星(GT/CA)_n 2碱基重复次数(3～52次)要远大于(GA/CT)_n 2碱基次数(3～27次)。获得的微卫星序列长度大小范围为129～601 bp,平均为286 bp。研究为进一步开展斑节对虾分子育种及资源评价分析提供了基础资料。