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.     

Serotonin induces ovarian maturation in giant freshwater prawn broodstock, Macrobrachium rosenbergii de Man

This study investigated the effects of serotonin (5-hydroxytryptamine or 5HT) on ovarian development in Macrobrachium rosenbergii de Man. Adult female prawns at the ovarian stage I (spent) were injected with 5HT at 1, 5, 10, 20 and 50 μg g^− 1 body weight (BW) intramuscularly on days 0, 5 and 10, and sacrificed on day 15. The doses as related to the effect could be categorized into three levels: low (1 and 5 μg g^− 1 BW of 5HT), medium (10 and 20 μg g^− 1 BW of 5HT) and high (50 μg g^− 1 BW of 5HT). The low-dose, especially at 1 μg g^− 1 BW, caused prawns to exhibit a significant increase in ovarian index (ovarian weight/body weight × 100) (5.79 ± 0.09%) as compared to the control (1.49%). The ovaries of most of these prawns could develop to stage IV (mature) and contained synchronously mature oocytes while most of the control ovaries remained at stage I and II (proliferative), and contained only oogonia to previtellogenic (Oc1, Oc2) and early vitellogenic oocytes (Oc3). The medium- and high-dose treated prawns exhibited ovaries that could reach stages III and IV and contained various types of oocytes of different maturity. Pretreatment with 5HT receptor antagonist, cyproheptadine (CYP), at 10 μg g^− 1 BW before 5HT injection significantly suppressed the effect of 5HT. Intramuscular injection of the 5HT-primed thoracic ganglion culture medium into CYP-pretreated prawns resulted in the increase of ovarian index about 5–6 times more than in the control, and in the groups injected with 5HT-primed media from muscle strip, eyestalk and brain. The ovaries of most prawn could develop up to stage IV and contained synchronously developed vitellogenic (Oc4) and mature oocytes (Oc5). These findings suggest that 5HT indirectly induces ovarian development and oocytes maturation in M. rosenbergii, probably via a putative ovarian stimulating factor released from the thoracic ganglia.     

The existence of gonadotropin-releasing hormone (GnRH) immunoreactivity in the ovary and the effects of GnRHs on the ovarian maturation in the black tiger shrimp Penaeus monodon

Immunocytochemical localization using antibodies against five isoforms of gonadotropin-releasing hormone (GnRH), namely, luteinizing hormone-releasing hormone (LHRH), salmon (s)GnRH, octopus (oct)GnRH, lamprey (l)GnRH-I, and lGnRH-III, showed that only lGnRH-I immunoreactivity (ir-lGnRH-I) was localized in follicular cells of proliferative, vitellogenic, and mature ovaries. The effects of exogenous GnRHs on the ovarian maturation cycle of Penaeus monodon were compared by treating female broodstocks with LHRH, sGnRH, and lGnRH-I. The cycle of ovarian maturation in both eyestalk-ablated and eyestalk-intact shrimp administered with the three isoforms of GnRH was significantly shorter than that of the control animals. Moreover, administrations of all GnRH isoforms showed similar numbers of spawned eggs and the percentage of successful fertilization as in the control animals. These findings suggest that GnRHs may be highly conserved peptides that play an important role in inducing the ovarian maturation in the shrimp.     

Effects of different feeding regimes on ovarian maturation and spawning of pond-reared giant tiger prawn in Thailand

Large pond-reared Penaeus monodon were used in three maturation and spawning trials lasting 60 days. One group of prawns was fed a fresh diet only; a second group was supplied a fresh diet combined with a formulated pelleted diet; a third group of prawns received the formulated diet only. Total maturation (N = 66 and 55) and spawning events (N = 63 and 48) were much greater for prawns receiving only the fresh diet, and the combined fresh with formulated diets, respectively. Prawns receiving only the formulated diet matured only 12 times, and spawned 11 times. Prawns fed a fresh diet produced significantly more eggs than those given a formulated diet alone. No significant differences were seen in eggs produced per spawning event, % fertilization, % hatch or % metamorphosis from egg to protozoea. Female prawns exhibited greater mortality than did males, while females on the fresh diet only had greater mortality than did other females, presumably due to greater handling and other stress factors. Female prawns on a combined diet had lower mortality than other females. The formulated diet used is suitable for supplementation, but not as a sole maturation food.     

Methyl farnesoate inhibition of late stage ovarian development and fecundity reduction in the black tiger prawn, Penaeus monodon

There is mounting evidence that the terpenoid hormone methyl farnesoate (MF) plays important roles in regulating reproductive processes in crustaceans. To gain further information on its roles and possible modes of action, MF was orally administered to ablated Penaeus monodon at a concentration of 5.5 μg per gram of diet, and a range of reproductive performance criteria measured. Results confirmed that MF plays a role in the reproductive process of this species. Specifically, under the conditions of this study, MF inhibited late stage ovary development and reduced fecundity in ablated prawns. The impact of the artificial diet (without additional MF) relative to a squid–mussel diet, was also assessed in this study and although it increased the quality of larvae produced, it also increased inhibition of late stage ovary development. Thus while the current study has increased our knowledge of MF by isolating an ovary developmental stage at which MF regulates reproduction in P. monodon, factors that determine the extent of its effect and whether it has a stimulatory or inhibitory effect, remain unknown. Until these factors are identified, the application of MF as a means of predictably manipulating egg production in captive prawns remains problematic.     

Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.     

中国南海海域斑节对虾群体与西印度洋、西太平洋群体种群遗传结构的比较分析

采用聚合酶链式反应(PCR)技术对海南三亚、深圳、湛江、北海4个斑节对虾群体共95个个体的延伸因子1-alpha 内含子序列进行了扩增,对扩增产物进行克隆转化,并将阳性克隆产物进行序列测定,最终获得了大小约216bp的可供分析的核苷酸序列。将获得的序列与从Genbank上下载的西太平洋、西印度洋序列进行比较分析。数据分析结果表明：北太平洋中国海域的基因多样性最低,西太平洋海域基因多样性水平最高;通过对遗传分化指数Fst的分析,发现西太平洋群体和西印度洋群体之间以及两者与北太平洋中国海域群体间遗传分化具有极显著性差异（P < 0.001）。UPGMA系统树显示,10个斑节对虾群体形成两大分支,一支由西太平洋群体和北太平洋中国海域群体组成,另一支由西印度洋群体单独组成;北太平洋中国海域群体中北海群体单独聚成一支。从序列差异的分析中得出,北太平洋中国海域群体与西印度洋群体之间的亲缘关系最远,与西太平洋群体之间的亲缘关系较近;中国海域内部各群体之间,北海群体与海南、深圳、湛江群体之间的亲缘关系较远,形成一个独特的地理种群。     

Heritability of reproductive traits and genetic correlations with growth in the black tiger prawn Penaeus monodon reared in tanks

Captive-reared broodstock of the black tiger prawn (Penaeus monodon) have exhibited poor reproductive performance limiting attempts to domesticate this species. The potential for improved reproductive performance was assessed by determining heritabilities of four measures of reproductive performance, their genetic correlations with each other and with growth rate and weight at age. Heritability estimates (h² ± S.E.) obtained from √ (days to spawn), √ (egg number), √ (nauplii number) and arcsin√ (proportion hatched) were 0.47 ± 0.15, 0.41 ± 0.18, 0.27 ± 0.16, and 0.18 ± 0.16, respectively. Estimates of genetic correlations between reproductive traits and weight at age, or growth rate were less than 0.5 except for √ (egg number) and weight at 54 weeks (0.93 ± 0.19) and √ (egg number) and 16–54-week growth (0.63 ± 0.29).     

Hemagglutinating activity and electrophoretic pattern of hemolymph serum proteins of Penaeus monodon and Macrobrachium rosenbergii to white spot syndrome virus injections

The giant fresh water prawn Macrobrachium rosenbergii is known to be highly tolerant to white spot syndrome virus (WSSV) infections when compared to the widely cultured marine tiger shrimp Penaeus monodon. At present, the exact mechanism of tolerance by M. rosenbergii to WSSV is not known. In this study, we attempt to study the effect of WSSV injections on the hemagglutination activity of the hemolymph serum of both P. monodon and M. rosenbergii and look for changes if any, on their hemolymph serum protein electrophoretic patterns. Our results show that M. rosenbergii had significantly (p < 0.05) higher hemagglutinating activity against mouse erythrocytes when compared to P. monodon. As the infection progressed to 48 h there was a further increase (p < 0.05) in the hemagglutination activity in M. rosenbergii, while it decreased in P. monodon. 12% SDS-PAGE analysis of the hemolymph serum of M. rosenbergii infected with WSSV did not show any new protein bands, whereas few bands with decreased intensity was observed in moribund P. monodon where the hemagglutinating activity was also observed to be decreased. The results indicate that hemolymph hemagglutinin levels are modulated in crustaceans as a response to viral infections.     

日本囊对虾卵母细胞发育过程中组织蛋白酶C比活性测定

组织蛋白酶C是一种溶酶样半胱氨酸蛋白酶,近年我们报道了组织蛋白酶C的表达与日本囊对虾(Marsupenaeus japonicus) 卵母细胞最后成熟阶段密切相关,本研究对卵母细胞发育主要不同时相特别是卵母细胞最后生理成熟过程进行了连续观察,测定了组织蛋白酶C在卵母细胞发育过程中的比活性。结果表明,日本囊对虾卵母细胞发育可划分为9个时相,即卵原细胞时相、核仁时相、周边核仁时相、初级卵黄发生时相、次级卵黄发生时相、皮质棒早期时相、皮质棒中期时相、皮质棒晚期时相、排卵期时相,生发泡的破裂是在排卵之前就已经发生;免疫印迹实验结果显示组织蛋白酶C蛋白只在卵母细胞最后成熟阶段即皮质棒发生时期表达,但酶活性测定结果表明组织蛋白酶C比活力在即皮质棒发生前后没有变化,酶活性较低,以上数据提示组织蛋白酶C可能与受精时皮质棒的释放以及受精卵外胶膜的形成有关。     

Inheritance mode of microsatellite DNA markers and their use for kinship estimation in kuruma prawn Penaeus japonicus

The allelic inheritance mode of microsatellite DNA markers was examined using seven copulated wild females and their offspring. Five microsatellite loci, CSPJ002 *, CSPJ010 *, CSPJ012 *, CSPJ014 *, and CSPJ015 *, were used in the study. At almost all family/locus combinations, one sire was determined and distributions of genotypes in offspring were consistent with the Mendelian segregation ratio. Distributions of genotypes were consistent with the ratio after assuming a null allele at some loci. Consequently, the alleles of CSPJ002 * and CSPJ012 * were inherited following the Mendelian inheritance mode in every family; however, the null allele was expected in CSPJ010 *, CSPJ014 *, and CSPJ015 * in some families. Thus, these loci should be used carefully in population genetic analysis, but siblings could be detected in the dendrograms based on unweighted pair-group method using arithmetic averages (UPMGA).     

南美白对虾的性腺发育、交配、产卵和受精

对池养南美白对虾 (Penaeusvannamei)亲虾进行人工促熟 ,并使之交尾、产卵、受精。实验在上海金山区杭州湾河口低盐地区一套自行设计的水循环系统中进行 ,育苗用水通过兑配使盐度为 2 7～ 2 8。育苗期间基本不换水和移动亲虾。亲虾 2 0 5尾 ,雌雄比约 1∶1。雌虾切除眼柄后 3～ 4d ,性腺即开始发育 ,6～ 7d产卵 ,但未受精。手术后约 30d ,亲虾出现追尾现象、随之发生交配并出现受精卵。实验进行了约 6 0d ,共收集虾卵 4 5 6 .7万粒 ,其中受精卵出现后所产卵为 137万粒 ,通过设置在水循环系统内的集卵网袋 ,可收集到 95 %以上的卵 ,共获受精卵 6 1万粒 ,平均日受精率为 4 4 .5 % ,无节幼体 2 5 .1万尾 ,受精卵的平均孵化率为 4 1.2 % ,最高达 87.5 %。文中还重点讨论了雌雄亲虾性腺发育的特点 ,产卵交配规律 ,以及亲虾规格大小、饵料种类、水质状况对南美白对虾交配和受精的影响     

The effects of ozone and probiotics on the survival of black tiger shrimp (Penaeus monodon)

Total ozone production (TOP) from an ozonator, residual ozone concentration (ROC) in water, and the effects of ozone with or without probiotic supplemented feeds on bacterial growth, and shrimp (Penaeus monodon) survival were investigated. Minimal effective ROC to inhibit 3 log units of Vibrio harveyi D331 for 6 h and 2 log units of Bacillus S11 for 9 h was 0.38 g O₃/l of ROC from 5-min ozonation. Shrimp postlarvae exposed to 0.34–0.50 mg O₃/l ROC (8-h ozonation) caused loss of balance, immobility and destruction of gill lamellar epithelium. In vivo treatment of juvenile P. monodon reared on probiotic feed for 1 month revealed that 0.35 mg O₃/l ROC (30-min ozonation) effectively inhibited 3 log units of V. harveyi D331 for 24 h. At this ROC dosage, there was no effect either on shrimp or on intestinal probiotics (Bacillus S11). Shrimp survival from probiotic treatment, coupling with ozonation, increased significantly (P≤0.05) compared with controls.     

金刚虾养殖现状及存在的问题分析

为研究亚硝态氮(NO₂-N)对斑节对虾(非洲群体)氧化应激、能量代谢和渗透平衡的影响,实验选取体长(3.0±0.5) cm的斑节对虾,设置0 (对照组)、5、10和15 mg/L(3个胁迫组)的亚硝态氮浓度梯度,进行了为期72 h的急性胁迫。实验结果显示,肝胰腺氧化应激因子活性(或含量)随着胁迫时间发展而变化。超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)以及一氧化氮合酶(NOS)活性先升高后下降,24 h,SOD和GSH-Px、NOS活性达到最大值,10 mg/L胁迫组的SOD、GSH-Px活性和15 mg/L胁迫组的NOS活性显著高于其他3组;丙二醛(MDA)含量变化相反,24 h胁迫组的MDA含量最小,其中10 mg/L胁迫组显著低于其他3组;48 h,CAT活性达到最大值,10 mg/L胁迫组显著高于其他3组;一氧化氮(NO)含量随着胁迫时间的延长而逐渐升高,72 h,NO含量最高,15 mg/L胁迫组显著高于其他3组。血清能量代谢指标中脂肪酶(LPS)活性先上升后下降,48 h,LPS活性达到最大值,其中10 mg/L胁迫组显著高于其他3组;而己糖激酶(HK)、磷酸果糖激酶(PFK)、丙酮酸激酶(PK)活性和甘油三酯(TG)含量明显下降,72 h,HK、PFK、PK活性和TG含量达到最小值,15 mg/L胁迫组显著低于其他3组。鳃丝中渗透调节参数——Na⁺、K⁺-ATP酶(Na⁺、K⁺-ATPase)活性在胁迫过程中始终下降,经过72 h的亚硝态氮胁迫,Na⁺、K⁺-ATPase活性达到最小值,15 mg/L胁迫组显著低于其他3组。研究表明,斑节对虾可通过提高抗氧化能力,增强对脂质吸收和利用,调节原有渗透平衡来应对亚硝态氮胁迫,这种作用的发挥具有一定的时间和浓度限制。本实验将为进一步探索亚硝态氮胁迫的分子机制提供数据支持。     

中华绒螯蟹卵巢快速发育期内脂类积累以及对抱卵的影响

本文报道中华绒螯蟹卵巢快速发育期内（９月初至１２月初）脂类积累及对抱卵的影响。结果表明：在这一时期,卵巢重量和脂肪含量都迅速增加,增加的脂肪中,中性脂（ＮＬ）主要是甘油三酯和胆固醇磷脂（ＰＬ）主要是卵磷脂和脑膦脂,且ＮＬ或ＰＬ在总脂的比值比较稳定;无论是ＮＬ还是ＰＬ,ω３系列的多不饱和脂肪（主要是Ｃ２０：５和Ｃ２２：６）的百分组成熟显著增加,而ω６系列的（主要是Ｃ２０：４）百分组成则显著降低;不能     

Differentiation of gonad maturation in sibling precocious males of the sea trout (Salmo trutta m. trutta L.) in their first year of life

Age at maturity and dynamics of gonadal cycle vary among individuals in a standard population; the extent of that variability in a group of siblings is not known for any species. Therefore, the aim of the study was to find out how the differentiation of gonad development varies in a group of sibling fish. The study involved 176 seven-month-old sea trout parr (mean fork length of 11 cm) collected at a single time point during their first spawning season (late October) from the brook of Pniewa in Western Pomerania (Poland). Gonad maturation was determined from histological analysis which was interpreted according to specially developed criteria.

The stage and forms of gonad maturation were diverse in the sibling group studied. Gonads of 52.8% males were not maturing (Class I), with only spermatogonia A present. The remaining males (47.2%) had gonads differing widely in the development, from beginning of maturation to completion of the reproductive cycle (Classes from II to VIII). In addition to the classes described, some males were observed to feature attempted maturation. It is difficult to provide an unequivocal estimate of the proportion of precocious males (mature at a young age, prior to the first descent to the sea) in the first year of life because of difficulties in categorizing males in Class II. It was predicted that 32.4% of the population would complete the cycle during the first spawning season. The body length and weight of the maturing males was significantly smaller (P < 0.05) than that of the non-maturing males.

Three forms of gonad maturation were identified. Gonads of a large group of precocious males (70%) developed incompletely (only some germ cell developed during the reproductive cycle), some of those gonads assumed the form of attempted maturation and partial maturation. Testes of the remaining 30% of the males underwent complete maturation. Gonads that undergo attempted maturation do not spermiate.

It is only from microscope-aided analysis of the testes that the class of gonad development can be accurately evaluated and the extent of precocious maturation assessed, particularly in fish maturing for the first time. It is impossible to correctly identify maturing males of the underyearling sea trout on the basis of morphological examination of gonad appearance, abdominal massage to detect spermiation or gonadosomatic index value.     

Quantifying the effects of bycatch reduction devices in Queensland''s (Australia) shallow water eastern king prawn (Penaeus plebejus) trawl fishery

This study presents results from an experimental 10-day research charter that was designed to quantify the effects of (a) a turtle excluder device (TED), (b) a radial escape section bycatch reduction device (BRD) and (c) both devices together, on bycatch and prawn catch rates in the Queensland shallow water eastern king prawn (Penaeus plebejus) trawl fishery. The bycatch was comprised of 250 taxa, mainly gurnards, whiting, lizard fish, flathead, dragonets, portunid crabs, turretfish and flounders. The observed mean catch rates of bycatch and marketable eastern king prawns from the standard trawl net (i.e., net with no TED or BRD) used during the charter were 11.06 kg/hectare (ha⁻¹) (S.E. 0.90) swept by the trawl gear and 0.94 kg ha⁻¹, respectively. For the range of depths sampled (20.1–90.7 m), bycatch rates declined significantly at a rate of 0.14 kg ha⁻¹ for every 1 m increase in depth, while prawn catch rates were unaffected. When both the TED and radial escape section BRD were used together, the bycatch rate declined by 24% compared to a standard net, but at a 20% reduction in marketable prawn catch rate. The largest reductions were achieved for stout whiting Sillago robusta (57% reduction) and yellowtail scad Trachurus novaezelandiae (32% reduction). Multidimensional scaling and analysis of similarities revealed that bycatch assemblages differed significantly between depths and latitude, but not between the different combinations of bycatch reduction devices. Despite the lowered prawn catch rates, the reduced bycatch rates are promising, particularly for S. robusta, which is targeted in another fishery. Prawn trawl operators are not permitted to retain S. robusta and the devices examined herein offer the potential to significantly reduce the incidental fishing mortality that this species experiences.     

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).     

Structure and function of antimicrobial peptide penaeidin-5 from the black tiger shrimp Penaeus monodon

The gene for penaeidin-5, an antimicrobial peptide comprising 55 amino acids, was isolated from the hemocyte of black tiger shrimp (Penaeus monodon). RT-PCR expression tests revealed that penaeidin-5 was produced in hemocytes, gills, the intestine and muscle. Western blot analysis confirmed the panaeidin-5 was aboundantin hemocytes, the intestine and hemolymph. Immunohistochemistry revealedpenaeidin-5 in the cuticle and gills that are considered primary defense barriers. The deduced amino acid sequence of penaeidin-5 included a proline-rich N-terminal domain and a carboxyl-domain that contained six cysteine residues. Circular dichrosim analysis revealed an -helix in its secondary structure and the predicted 3D structure indicated two-disulfide bridges in the -helix. Based on the sequence of penaeidin-5 peptide cDNA, synthetic penaeidin-5 was prepared to carry out functional tests. The synthetic peptide had efficient bacteriostatic and bactericidal activity against Aerococcus viridans, and also inhibited the growth of two filamentous fungi, Fusarium pisi and Fusarium oxysporum. To measure penaeidin-5 in vivo, black tiger shrimp were challenged with Vibrio alginolyticus and A. viridans. At 3 h post-challenge, penaeidin-5 was induced and bacterial numbers decreased significantly by 12 h and 24 h.