Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.     

Culturing freshwater pearl mussel Margaritifera margaritifera: a breakthrough in the conservation of an endangered species

• 1. The freshwater pearl mussel Margaritifera margaritifera L. is globally endangered and is threatened by commercial exploitation, pollution and habitat loss throughout its range. Captive breeding would be a valuable tool in enhancing the status of M. margaritifera in the UK.
• 2. We have developed a semi‐natural system for successfully infecting juvenile brown trout with glochidial M. margaritifera, and culturing juvenile mussels in experimental tanks where glochidial M. margaritifera can excyst from fish gills and settle into sediment.
• 3. Infected fish had less than 1% mortality. Levels of infection varied among fish. Two yearly cohorts of juvenile M. margaritifera were identified from samples of sediment taken from each experimental tank. Individuals range in size from 1.4 mm (2000 cohort) to >3 mm in length (1999 cohort).
• 4. The number of juvenile M. margaritifera present in the two experimental tanks are estimated to be between 3600 (tank A) and 0 (tank B) for the putative 1999 cohort and between 6000 (tank A) and 13 000 (tank B) for the putative 2000 cohort.
• 5. This pioneering method for large‐scale cultivation of juvenile M. margaritifera is intermediate between the release of infected fish into rivers and the intensive cultivation systems developed in continental Europe and the USA for other species of unionid. This is the first time that large numbers of M. margaritifera have been cultured and represents a significant breakthrough in the conservation of this globally endangered Red Data List species. The method is straightforward and is most cost‐effective when undertaken alongside established hatchery processes.

Copyright © 2006 John Wiley & Sons, Ltd.     

三江平原湿地开垦对地表水中可溶性铁分布特征的影响

选取三江平原湿地开垦前后具有代表性的地表水类型,以空间换时间的方法对比分析了湿地开垦对地表水中可溶性铁（WSIFe）含量与形态分布的影响。结果表明,在田块尺度上,湿地地表积水、排水中WSIFe的含量均值分别为2.20、1.75mg·L^-1;开垦为大豆田后地表无积水,排水中降为1.34mg·L^-1;开垦为水田后分别增加为2.76、2.38mg·L^-1,这与使用铁含量较高的地下水灌溉有关。在区域尺度上,虽然湿地开垦导致沼泽性河流中WSIFe含量降低,但目前含量（均值为0.66mg·L^-1）仍高于黑龙江（0.34mg·L^-1）,是黑龙江WSIFe的重要来源;农田排水干渠中WSIFe含量均值为0.42mg·L^-1,也是黑龙江WSIFe的来源之一,但其贡献量低于沼泽性河流;湿地开垦减少了地表水中WSIFe向黑龙江的输出。湿地开垦后地表水中Fe^3＋的相对含量在田块尺度上出现增加,在区域尺度上有所降低,湿地开垦前后地表水中WSIFe的主要存在形态均为Fe^3＋。无论是田块尺度还是区域尺度,湿地开垦后地表水中WSIFe含量的季节变化都与灌溉活动有关。     

中国对虾育苗时间与温度的关系及其应用

根据多年的调研和试验资料，利用非线性积温模型，模拟了对虾幼体各发育阶段所需天数与水温的关系。在此基础上研究确定了亲虾产卵孵化期的气候决策模式；并应用模式计算出了我市亲虾产卵孵化的适宜时段。为亲虾适宜产卵孵化期的决策提供了科学方法。     

Chemical and Thermal Properties of Freshwater Prawn (Macrobrachium rosenbergii) Meat

Chemical compositions and thermal properties of cultured freshwater prawn meat (FPM) were studied. FPM contained 83.2% protein (dry basis), 62.7% of which was myofibrillar protein. Pepsin-soluble collagen (PSC) and insoluble collagen (ISC) contents were 0.63 and 0.32%, respectively. Both collagens were similar to type V collagen from porcine placenta. Glutamic acid/glutamine, arginine, aspartic acid/?asparagine, and lysine were abundant amino acids in FPM. Glycine, proline, hydroxyproline, and aspartic acid/?asparagine were predominant in both collagens. FPM exhibited thermal transition temperatures (T_max) of 48.3 and 64.7°C, whereas T_max of PSC and ISC were 43.0 and 46.0°C, respectively. Textural changes in FPM during post-mortem storage on ice are plausibly dependent upon its compositional and thermal properties.     

Median Lethal Concentration (LC50) of Piscicidal Plants and Their Utilization in Aquaculture

Aqueous and ethanol extracts of Acacia pennata (bark), Catunaregam uliginosa (fruit), Diospyros lenceofolia (fruit), and Sapindus mukorossi (fruit) were evaluated as piscicides on the freshwater fish Danio dangila, Danio rerio, and Puntius shalynius, and the air-breathing Heteropneustes fossilis. Catunaregam uliginosa (fruit) possessed relatively higher piscicidal potency followed by Sapindus mukorossi (fruit), Acacia pennata (bark), and lastly Diospyros lanceofolia (fruit). Catunaregam uliginosa can thus be used for eradicating unwanted wild fishes in a short time span at low concentrations so that the health of aquatic environment is maintained.     

Pathogenicity of halophilic Vibrio harveyi in giant freshwater prawns (Macrobrachium rosenbergii De Man)

In this study, the phenotypic and pathogenic properties in freshwater host were characterized in 14 strains of halophilic Vibrio harveyi isolated from infected marine black tiger shrimp, Penaeus monodon. The lysogenic phenotype was assayed via prophage excision and mitomycin C induction. The bacteria were grouped into two types, corresponding to lysogenic and non‐lysogenic strains. The pathogenicity was determined via direct injection of bacterial cultures into post‐larval juvenile giant freshwater prawns, Macrobrachium rosenbergii De Man. All of the infected prawns showed similar symptoms and inflamed hepatopancreas. The V. harveyi isolates derived from the first‐injected infected prawns were re‐isolated and re‐injected into healthy giant freshwater prawns, in which they retained similar infectivity. Both lysogenic and non‐lysogenic Vibrio spp. showed identical virulence associated with 100% mortality within one day post‐injection. TEM micrographs showed hepatopancreatic nuclear deformation and lipid breakdown caused by lysogenic γ‐hemolytic VL19 and non‐lysogenic β‐hemolytic V33. However, the V33 strain was associated with severely disrupted mitochondria. None of the V. harveyi strains was able to produce a biofilm. Together, our findings indicate that the lysogenic and non‐lysogenic halophilic V. harveyi isolated from marine shrimps may use different virulence factors that are responsible for their pathogenicity in freshwater prawns.     

The On-Farm Trials of the Rotational Prawn-Rice Farming in a Semi-Deep Water Area

ABSTRACT

The on-farm trials of rotational rice-prawn farming in a semi-deep water area in the Mekong Delta of Vietnam was carried out in six rice fields (0.7–1.0 ha) to evaluate the production and economic efficiency of using low and high cost feed on prawn culture in paddies at different densities. Two stocking densities of 4 and 5 PL/m² were investigated. Two treatments of feed types (pellets only or a combination of pellets, trash fish, and snail meat) were applied at stocking 4 PL/m². At the treatment of 5 PL/m², prawns were fed a combination of pellets, trash fish, and snail meat. Prawns were stocked in the mid-April and harvested in mid‐November before the next dry season rice crop. Rice farming was started 3 to 5 days after prawn harvesting. By cull harvesting during the culture period, final mean weights of prawn ranged from 47.2 to 57.2 g/prawn and the male:female ratio at harvesting was 2.5:1.0. The prawn yield of treatment 5 PL/m² was highest (630 ± 22 kg/ha). Net profits in treatments using the combination of pellets, trash fish, and snail meat were 861 ± 193 US$/ha to 1,019 ± 25 US$/ha for the prawn crop and 1,393 ± 71 US$/ha to 1,576 ± 180 US$/ha for the whole system (prawn crop + dry rice crop) and significantly higher than in treatment using pellet only (P < 0.05). Prawns fed on pellets or a combination of pellets and snail meat both offer similar results in terms of production and economics. The dry rice crop offered high cost benefit ratio (2.29–2.33) with low operating cost (414–434 USD/ha) and it made a better use in sustainable rice fields through the rotational rice-prawn system.