Dietary arginine requirement of juvenile kuruma shrimp Marsupenaeus japonicus (Bate)

The requirements for essential amino acids by kuruma shrimp, Marsupenaeus japonicus (Bate) have not yet been determined using the dose–response method due to poor utilization of crystalline amino acids (CAA). In a previous study, we produced coated CAA that could be used to retard leaching losses. Therefore, to determine the arginine requirement of juvenile kuruma shrimp, a growth study was conducted using test diets containing six levels of coated arginine (from 1.20% to 3.19% of diet) for 42 days. Crystalline amino acids was supplemented to correspond to the amino acid pattern in the whole body protein of the kuruma shrimp except for arginine. To reduce leaching losses, CAA were coated with carboxymethylcellulose (CMC) and diets were further bound with CMC and k‐carrageenan. Test diets were fed to triplicate groups of juveniles (0.25±0.02 g) twice a day. Proximate composition and the amino acid composition of shrimp whole bodies were analysed at the end of the feeding trial. Percent body weight gain (WG), specific growth rate, feed intake, feed efficiency ratio and protein retention efficiency were significantly affected by the dietary arginine concentrations. Estimating the arginine requirement of kuruma shrimp using broken‐line analysis of percent WG resulted in a value of 2.66% of diet or 5.32% of protein. After correcting for leaching of arginine from the diets, the optimum dietary value of arginine for kuruma shrimp was a little lower than the estimated value.     

Comparison of defence ability against the white spot syndrome virus between Fenneropenaeus chinensis and Marsupenaeus japonicus

The defence ability of Fenneropenaeus chinensis and Marsupenaeus japonicus against the intrusion of the white spot syndrome virus (WSSV) was compared after injecting WSSV intramuscularly by recording cumulative mortality, diagnosing the virus and examining variations in immunological parameters including the total haemocyte counts (THCs), phagocytic percentage (PP), plasma protein concentration (PPC), phenoloxidase (PO) and nitric oxide synthase (NOS) activity. The results showed that the variations in the immunological parameters of F. chinensis and M. japonicus showed similar trends. The THCs of the two species decreased significantly postchallenge of WSSV. The virus was detected at 78 h in M. japonicus and at 42 h in F. chinensis after infection, which was in correlation with the accumulative mortality, and the variations in PO, PP, NOS and PPC in the two species. All shrimps of F. chinensis in the mortality experiment died within 66 h, much more quickly than M. japonicus, whose cumulative mortality reached 100% after 198 h. In conclusion, the immune system of M. japonicus has a stronger resistant ability to antagonize and endure the invasion of WSSV than that of F. chinensis.     

Effects of water-soluble and fat-soluble choline sources on the performances of juvenile kuruma shrimp, Marsupenaeus japonicus Bate

The present study was undertaken to clarify the effect and interaction of choline chloride (CC) and phosphatidylcholine (PC), the water‐ and fat‐soluble sources of choline, on the performances of kuruma shrimp, Marsupenaeus japonicus juveniles using as response variables such as percent weight gain (%WG), specific growth rate (SGR), survival (SR%), feed efficiency ratio (FER), protein efficiency ratio (PER), proximate composition, fatty acids profile and free choline and PC contents in the whole body of kuruma shrimp. A 40‐day feeding trial was conducted as a 2 × 3 factorial experiment with six test diets containing three levels of CC (0.3, 0.6 and 1.2 g kg^?1) and two levels of soybean PC (SPC; 2 and 4 g kg^?1). Both of the choline sources (CC and SPC) showed a significant interaction (P<0.05) regarding all the evaluated response variables except for the proximate and fatty acid contents. Interestingly, there was a positive effect (P<0.05) of CC observed with increasing its level at the low level of SPC and vice versa. However, the magnitudes of the beneficial effect of CC in terms of the evaluated parameters were changed with the different dietary levels of SPC. Also, the inclusion of higher levels of SPC (4 g kg^?1) or CC (0.6 and 1.2 g kg^?1) diminished the effect of the other source.     

Effects of supplementation of coated crystalline amino acids on growth performance and body composition of juvenile kuruma shrimp Marsupenaeus japonicus

Supplementation of crystalline amino acids (CAA) in shrimp diets is unsuccessful in most cases. In the present study, various pre‐coated CAA were developed to minimize the leaching losses from diets in water and assessed using kuruma shrimp Marsupenaeus japonicus juveniles. Experimental diets were supplemented with coated CAA or non‐coated CAA to simulate the dietary amino acid profile to that found in the whole body protein of shrimp. The four diets contained CAA coated with either carboxymethyl cellulose (CMC) (diet 1), zein (diet 2), k‐carrageenan (diet 3) or agar (diet 4), respectively. Another four diets contained CMC‐coated CAA with the addition of either zein (diet 5), casein‐gelatin (diet 6), zein and k‐carrageenan (diet 7), or casein‐gelatin and k‐carrageenan (diet 8), respectively. The 8 weeks feeding trial demonstrated that the diets containing coated CAA gave significantly higher weight gains than diets containing non‐coated CAA, with the highest value for the diet 6. The diets containing coated CAA also yielded higher feed conversion efficiency, protein efficiency ratio, and specific growth rate than diets containing non‐coated CAA. Leaching trials suggested that shrimp growth performance was improved partly due to the retardation or reduction of CAA leaching by coating them with specific binders.     

Short-term responses of adult kuruma shrimp Marsupenaeus japonicus (Bate) to environmental salinity: osmotic regulation, oxygen consumption and ammonia excretion

The effects of salinity on haemolymph osmolality, oxygen consumption and ammonia excretion were investigated in adult kuruma shrimp, Marsupenaeus japonicus (Bate), at salinities of 20, 25, 30 and 35 g L^?1. Haemolymph osmolality showed a positive linear relationship (r²=0.9854) with medium osmolality. The isosmotic point calculated from this relationship was 1039 mOsm kg^?1, which corresponds to a salinity of approximately 35 g L^?1. The slope of the regression equation was very high (0.81), suggesting that M. japonicus adults are poor osmoregulators compared with the adults of other penaeids and to conspecific young. The difference between haemolymph and medium osmolality (D_OP) was lowest at 35 g L^?1 and highest at 20 g L^?1. Thus, the minimum D_OP coincided with the isosmotic point of the shrimp. The respiration rate was significantly lower at 30 g L^?1 than at the other salinities. Ammonia excretion rates were inversely related with salinity and therefore were minimal at 35 g L^?1. The results of this study suggest that the optimum salinity for adult kuruma shrimp is around 30–35 g L^?1 and that even minor (e.g. 5 g L^?1) deviations from this optimum cause significant physiological changes. Further, the observed increases in oxygen consumption and ammonia excretion during exposure to low salinities, which indicate higher energy expenditure and amino acid catabolism for osmoregulation, respectively, suggest that the growth efficiency of M. japonicus adults may be severely compromised by hypohaline water inflow into the rearing ponds.     

Recent insights into host–pathogen interaction in white spot syndrome virus infected penaeid shrimp

Viral disease outbreaks are a major concern impeding the development of the shrimp aquaculture industry. The viral disease due to white spot syndrome virus (WSSV) observed in early 1990s still continues unabated affecting the shrimp farms and cause huge economic loss to the shrimp aquaculture industry. In the absence of effective therapeutics to control WSSV, it is important to understand viral pathogenesis and shrimp response to WSSV at the molecular level. Identification and molecular characterization of WSSV proteins and receptors may facilitate in designing and development of novel therapeutics and antiviral drugs that may inhibit viral replication. Investigations into host–pathogen interactions might give new insights to viral infectivity, tissue tropism and defence mechanism elicited in response to WSSV infection. However, due to the limited information on WSSV gene function and host immune response, the signalling pathways which are associated in shrimp pathogen interaction have also not been elucidated completely. In the present review, the focus is on those shrimp proteins and receptors that are potentially involved in virus infection or in the defence mechanism against WSSV. In addition, the major signalling pathways involved in the innate immune response and the role of apoptosis in host–pathogen interaction is discussed.     

Apoptosis does not play an important role in the resistance of 'immune' Penaeus japonicus against white spot syndrome virus

We previously demonstrated that kuruma shrimp, Penaeus japonicus, exposed to white spot syndrome virus (WSSV) became resistant ('immune' shrimp) to subsequent challenge with the virus. The present study investigated the role of apoptosis in the 'immune' shrimp during a secondary challenge with WSSV. When naive kuruma shrimp were intramuscularly injected with WSSV at a high or low dose, apoptosis was often detected by TUNEL assay in the lymphoid organ (LO), mainly in the early stage of the infection. A significantly higher incidence of apoptosis was observed in the LO of the shrimp injected with the high dose of WSSV (cumulative mortality: 100%) than in the shrimp injected with the low dose (cumulative mortality: 0%). When 'immune' and naive shrimp were injected with an equal dose of WSSV, the incidence of apoptosis was significantly lower in the 'immune' shrimp than in the naive shrimp. This difference is assumed to result from a substantial reduction of the virus by humoral neutralizing factor in the 'immune' shrimp. These results suggest that apoptosis is not a principal protective factor in 'immune' shrimp.     

Testing of a pond‐side molecular diagnostic tool for the detection of white spot syndrome virus in shrimp aquaculture

White spot disease in penaeid shrimp is caused by the white spot syndrome virus (WSSV). It is the most economically important disease of farmed warm‐water shrimp, causing extensive economic losses estimated from $8 to $15 billion since its emergence in the 1990s. Early diagnosis of disease is critical in the management of outbreaks and to avoid crop losses. Diagnosis of white spot disease is generally carried out in centralized laboratory settings using molecular biology approaches. However, this mode of testing can be expensive and time consuming, requiring laboratory equipment, highly trained laboratory personnel, dedicated laboratory space, and long‐distance transportation of samples from field to lab. In‐field diagnostics are gaining credence as tools for rapid and early animal disease detection, allowing diagnosticians and farmers to potentially manage disease outbreaks from the pond side. In the present study, we describe the development and application of a new in‐field point‐of‐need diagnostic test and platform for the diagnosis of WSSV in remote settings (shrimp farms). We report its performance in laboratory and field settings and compare it with current gold‐standard diagnostic approaches. We discuss the potential benefits (and barriers to uptake) of applying such testing in the global shrimp farming sector.     

Interactive effect of two methyl‐group sources on the growth and phospholipids’ content of kuruma shrimp Marsupenaeus japonicus Bate post‐larvae

Six micro‐bound diets were formulated to contain three levels of choline chloride (CC) (0.0, 0.6 and 1.2 g kg⁻¹) and 2 levels of methionine (Met) (0 and 15 g kg⁻¹). Soybean protein isolates (SPI) were used as the main protein source for its limited Met content. A significant (P < 0.05) interaction was determined between CC and Met on the survival (S %), weight gain (WG%), specific growth rate (SGR % day⁻¹), feed efficiency ratio (FER), protein efficiency ratio (PER) and phosphatidylcholine (PC), phoshphatidylethanolamine (PE) and Met contents of the whole body of shrimp. The shrimp group did not receive either supplemental CC or Met showed lower (P < 0.05) values of the above‐mentioned parameters than other shrimp groups fed with 0.6 and 1.2 g kg⁻¹ supplemental CC with or without Met supplementation. The present study showed that supplementation of 1.2 g kg⁻¹ CC in the diets could compensate shrimp post‐larvae with the needed methyl group when received Met‐deficient diets. The study also assumed that the biosynthesis of PC in the shrimp’s body can be achieved by the methylation of PE through the S‐adenosylmethionine (SAM) pathway and/or through the cytosine di‐phosphoryl (CDP) choline pathway directly from dietary choline.     

The incidence of suspected white spot syndrome virus in semi‐intensive and extensive shrimp farms in Bangladesh: implications for management

The study was conducted to assess key factors influencing suspected white spot syndrome virus (WSSV) disease and associated shrimp production and economic performance in three contrasting black tiger shrimp (Penaeus monodon) culture technologies promoted by the United States Agency for International Development funded Shrimp Quality Support Project (SQSP) in Bangladesh. A total of 350 traditional, 315 Modified Traditional Technology1 (MTT1), 36 MTT2 and 88 Closed System Technology (CST) farmers from 10 sub‐districts in three districts of Khulna division were surveyed following random sampling at the end of the project. Binomial probit regression analysis revealed that smaller newly constructed ponds (known locally as gher) were less susceptible to WSSV, provided aquatic weeds were controlled using chemicals. Removal of sludge from ghers also had a positive effect, irrespective of technology and location. It was also shown that stocking of screened shrimp postlarvae (PL) does not guarantee protection against WSSV (t = 1.39, P > 0.05). Higher shrimp production was obtained by farmers practicing CST, followed by those operating MTTs and traditional technology respectively. Farmers who adopted CST also gained higher profitability followed by those operating MTT1, MTT2 and traditional technology.     

Effects of dietary astaxanthin supplementation on survival,growth and stress resistance in larval and post‐larval kuruma shrimp,Marsupenaeus japonicus

Two trials were conducted to evaluate the effects of chemically synthesized astaxanthin (Ax) on growth performance, survival, and stress resistance in larval and post‐larval kuruma shrimp (Marsupenaeus japonicus). Six dietary levels of Ax (0, 50, 100, 200, 400 and 800 mg/kg diet) were added to a baseline diet. As a first study, an 8‐day feeding trial was conducted on larval kuruma shrimp, with results showing that larvae‐fed diets supplemented with different Ax levels exhibited better performance during developmental and metamorphosis to postlarvae. Broken‐line regression analysis indicated that the optimal levels for growth and stress resistance of larvae were 168.9 mg/kg and 82.1 mg/kg diet, respectively. Second, a 30‐day feeding trial was conducted on post‐larval shrimp. Supplementation with 100 and 200 mg/kg Ax yielded significantly higher final body weight, body weight gain and specific growth rate than that in a control group. A cumulative mortality index for osmotic stress revealed significantly better performance in a group fed 200 mg/kg supplementation than in controls. Broken‐line regression analysis indicated that the optimal levels for growth and stress resistance of postlarvae were 108.7 mg/kg and 178.1 mg/kg diet, respectively. Based on these results, we suggest that carefully dosed Ax supplementation is a beneficial nutritional strategy for the early developmental stages of kuruma shrimp.     

对虾白斑综合征病毒在螯虾动物模型的感染特性

应用克氏原螯虾作为动物模型研究对虾白斑综合征病毒（ＷＳＳＶ）的感染增殖特性，涉及感染温度，感染途径，继发感染，半数致死量（ＬＤ５０），免疫保护及保存期等，结果显示，２２－２５℃时，接种ＷＳＳＶ的螯虾一般于２－７d内死亡，温度升高对病毒增殖影响不显著，３０－３２℃时，平均死亡时间２－６d，温度降低对病毒增殖影响较显著，１５－１９℃时，接种螯虾平均２－１０d内死亡，８－１０℃时，平均死亡时间３－６d，感染途径分别用腹节肌肉，腹节皮下洲射及口服，均能使螯虾感染发病，而浸泡方式不能使螯虾发病，细菌分离和细菌定量结果表明，寄考于螯虾心脏，肝胰腺内的阴沟肠杆菌在感染后期大量增殖，菌量分别是正常螯虾的２５和３０倍，形成继发感染，用螯虾心脏，肝胰腺内的阴肠杆菌在感染后期大量增殖，菌量分别是政党螯虾的２５和３０倍，形成继发梁。用螯虾测定ＷＳＳＶ的ＬＤ５０为１０^-6.5.mL^-1种毒液，将病毒５６℃，３０min灭活后免疫螯虾，不能使螯虾形成免疫保护，ＷＳＳＶ匀浆液－３０℃冻存１年后失活，而－３０℃冻存于螯虾体内ＷＳＳＶ保存１年后仍有活力。     

A synthetic antibacterial peptide from Mytilus galloprovincialis reduces mortality due to white spot syndrome virus in palaemonid shrimp

White spot syndrome virus (WSSV) isolated from Penaeus monodon was found to be highly infective for the western Mediterranean shrimp, Palaemon sp. Using polymerase chain reaction (PCR), it was demonstrated that such shrimp are not naturally carriers of WSSV. Following challenge with virus, mortality reached 100% 3.5-4 days after injection at 22 degrees C. Incubation of infected shrimp at 10 degrees C totally suppressed the mortality which rapidly developed when shrimp were returned to 18 or 22 degrees C. Preincubation of WSSV with mature synthetic mytilin significantly reduced shrimp mortality with a 50% efficient dose of about 5 microM. Survival of shrimp was not due to the development of an active mechanism of defence as re-injection of WSSV produced the same mortality pattern. Mortality was probably due to WSSV replication as dot blot failed to detect viral DNA in the injection sample but was positive 1 day post-injection. Protection by mytilin was by interaction at the virus level, preventing replication as no WSSV nucleic acid was detected by PCR even after 7 days in shrimp injected with WSSV preincubated with 10 or 50 microM mytilin.     

The effects of sediment redox potential and stocking density on Pacific white shrimp Litopenaeus vannamei production performance and white spot syndrome virus resistance

Redox potential represents the intensity of anaerobic condition in the pond sediment, which may affect the dominant microbial transformations of substances, the toxins production, mineral solubility, as well as the water quality in the sediment–water interface inhabited by the shrimp. This study evaluates the effect of sediment redox potential in conjunction with stocking density on shrimp production performance, immune response and resistance against white spot syndrome virus (WSSV) infection. A completely randomized two factors experimental design was applied with three different sediment redox potential, i.e. ?65, ?108 and ?06 mV, and two shrimp densities, i.e. low (60 shrimp m^?2) and high (120 shrimp m^?2). Shrimp juveniles with an initial mean body weight of 5.32 ± 0.22 g were maintained in semi‐outdoor fibre tanks (270 L in capacity) for 35 days of experimental periods. At the bottom of each tank, 5‐cm deep soil substrate with different redox potential was added according to the treatments. The survival and biomass production were significantly reduced at ?206 mV sediment redox potential, regardless of stocking density. Highly negative sediment redox potential (?206 mV) and higher stocking density significantly reduced total haemocyte counts and phenoloxydase activity, and shrimp resistance to WSSV infection. We recommend to maintain the redox potential of pond sediment at a level of more than ?206 mV.