Maturation of rhesus monkey oocytes in chemically defined culture media and their functional assessment by IVF and embryo development

This study compared success of in-vitro maturation of rhesus monkey oocytes in protein-free versus serum-containing culture systems, assessed by embryo development subsequent to IVF. Four media were tested: (i) modified Connaught Medical Research Laborato

Influence of selected Indian immunostimulant herbs against white spot syndrome virus (WSSV) infection in black tiger shrimp, Penaeus monodon with reference to haematological, biochemical and immunological changes

Immunostimulants are the substances, which enhance the non-specific defence mechanism and provide resistance against the invading pathogenic micro-organism. In order to increase the immunity of shrimps against the WSSV, the methanolic extracts of five different herbal medicinal plants like Cyanodon dactylon, Aegle marmelos, Tinospora cordifolia, Picrorhiza kurooa and Eclipta alba were selected and mixed thoroughly in equal proportion. The mixed extract was supplemented with various concentrations viz. 100 (A), 200 (B), 400 (C), and 800 (D) mg kg(-1) through artificial diets individually. The prepared diets (A-D) were fed individually to WSSV free healthy shrimp Penaeus monodon with an average weight of 8.0 +/- 0.5 g for 25 days. Control diet (E), devoid of herbal extract was also fed to shrimps simultaneously. After 25 days of feeding experiment, the shrimps were challenged with WSSV, which were isolated and propagated from the infected crustaceans. The shrimps succumbed to death within 7 days when fed on no herbal immunostimulant diet (E). Among the different concentrations of herbal immunostimulant supplemented diets, the shrimps fed on diet D (800 mg kg(-1)) significantly (P < 0.0001) had more survival (74%) and reduction in the viral load. Also the better performance of haematological, biochemical and immunological parameters was found in the immunostimulant incorporated diets fed shrimps. The present work revealed that the application of herbal immunostimulants will be effective against shrimp viral pathogenesis and they can be recommended for shrimp culture. (c) 2006 Published by Elsevier Ltd.

Effects of different light sources and illumination methods on growth and body color of shrimp Litopenaeus vannamei

Shrimps Litopenaeus vannamei with initial body weight of 2.108 +/- 0.036 g were sampled for specific growth rates (SGR) and body color measurements for 50 days under different light sources (incandescent lamp, IL; cool-white fluorescent lamp, FL; metal halide lamp, MHL; and control without lamp) and different illumination methods (illumination only in day, IOD, and illumination day and night, IDN). Body color of L. vannamei was measured according to the free astaxanthin concentration (FAC) of shrimp. The SGR, food intake (FI), feed conversion efficiency (FCE) and FAC of shrimps showed significant differences among the experimental treatment groups (P < 0.05). Maximum and minimum SGR occurred under IOD by MHL and IDN by FL, respectively (difference 56.34%). The FI of shrimp for the control group did not rank lowest among treatments, confirming that shrimp primarily use scent, not vision, to search for food. FI and FCE of shrimps were both the lowest among treatment groups under IDN by FL and growth was slow, thus FL is not a preferred light source for shrimp culture. Under IOD by MHL, shrimps had the highest FCE and the third highest FI among treatment groups ensuring rapid growth. FAC of shrimp were about 3.31 +/- 0.20 mg/kg. When under IOD by MHL and IDN by FL, FAC was significantly higher than the other treatments (P < 0.05). To summarize, when illuminated by MHL, L. vannamei had not only vivid body color due to high astaxanthin concentration but also rapid growth. Therefore, MHL is an appropriate indoor light source for shrimp super-intensive culture. SGR of shrimp was in significantly negative correlation to FAC of shrimp (P < 0.05). Thus, when FAC increased, SGR did not always follow, suggesting that the purpose of astaxanthin accumulation was not for growth promotion but for protection against intense light. (c) 2005 Elsevier B.V. All rights reserved.

Protective immunity induced by CpG ODNs against white spot syndrome virus (WSSV) via intermediation of virus replication indirectly in Litopenaeus vannamei

The worldwide shrimp culture is beset with diseases mainly caused by white spot syndrome virus (WSSV) and suffered huge economic losses, which bring out an urgent need to develop the novel strategies to better protect shrimps against WSSV. In the present study, CpG-rich plasmid pUC57-CpG, plasmid pUC57 and PBS were employed to pretreat shrimps comparatively to evaluate the protective effects of CpG ODNs on shrimps against WSSV. The survival rates, WSSV copy numbers, and antiviral associated factors (Dicer, Argonaute, STAT and ROS) were detected in Litopenaeus vannamei. There were higher survival proportion, lower WSSV copy numbers, and higher mRNA expression of Dicer and STAT in pUC57-CpG-pretreatment shrimps than those in pUC57- and PBS-pretreatment shrimps after WSSV infection. The Argonaute mRNA expression in pUC57-CpG-, pUC57- and PBS-pretreatment shrimps after WSSV infection was significantly higher than that of shrimps post PBS stimulation on the first day. The ROS levels in pUC57-CpG-pretreatment shrimps post secondary stimulation of PBS were significantly higher than those post WSSV infection on the first day. These results together demonstrated that pUC57-CpG induced partial protective immunity in shrimps against WSSV via intermediation of virus replication indirectly and could be used as a potential candidate in the development of therapeutic agents for disease control of WSSV in L. vannamei. (C) 2009 Elsevier Ltd. All rights reserved.

Production of astaxanthin from Haematococcus in open pond by two-stage growth one-step process

We have developed a two-stage growth one-step process for cultivation of Haematococcus using a self-designed system that mimics an open pond in the natural environment. The characteristics of this process are green vegetative cell growth and cysts transformation and pigment accumulation that proceed spontaneously and successively in one open photobioreactor. Four strains of Haematococcus (H. pluvialis 26; H. pluvialis 30; H. pluvialis 34; H. pluvialis WZ) were cultured in this imitation system for a duration of 12 days. The changes in cell density and medium pH were closely monitored, and the astaxanthin content and yield of the four Haematococcus strains were measured at the end of 12 days of cultivation. Two of the strains, H. pluvialis 26 and H. pluvialis WZ, were selected as strains suitable for mass culture, resulting in the astaxanthin yield of 51.06 and 40.25 mg L-1 which are equivalent to 2.79 and 2.50% of their dry biomass respectively. Based on the laboratory work, 6 batch cultures of H. pluvialis WZ were conducted successfully to produce astaxanthin in two 100 m(2) open race-way pond by two-stage growth one-step process. The astaxanthin content ranged from 1.61 to 2.48 g 100 g(-1) dry wt., with average astaxanthin content of 2.10 g 100 g(-1) dry wt. Compared with the one-stage production of astaxanthin based on continuous culture, the superiority of our process is that it can accumulate much more astaxanthin in red cysts. Compared with two-stage production of astaxanthin, the advantage of our process is that it does not need to divide the production process into two parts using two bioreactors. The presented work demonstrates the feasibility for producing astaxanthin from Haematococcus using a two-stage growth one-step process in open pond, culture systems that have been successfully used for Spirulina and Chlorella mass culture. The future of Haematococcus astaxanthin production has been also discussed. (C) 2009 Elsevier B.V. All rights reserved.

Growth of Gracilaria lemaneiformis under different cultivation conditions and its effects on nutrient removal in Chinese coastal waters

Gracilaria lemaneiformis (Bory) Daws has been extensively cultivated as a source of commercial agar and the ecomaterials in Shenao Bay, Guangdong Province, Jiaozhou Bay, Shandong Province and other waters in China. This paper examines the in situ suspended farming of G. lemaneiformis using raft cultivation under different conditions and its effects on nutrient removal in the laboratory. The results showed that cultivated Gracilaria grew well in both Shenao Bay and Jiaozhou Bay. The biomass of Gracilaria increased from 50 to 775 g m(-1) (fresh weight) during 28 days, with special growth rate (SPG) 13.9% d(-1) under horizontal cultivation in Jiaozhou Bay. Light, temperature, nutrient supply, as well as cultivation treatments such as initial density, and depth of suspension seaweed were important to the growth of Gracilaria. The highest biomass production was observed in the horizontal culture condition (0.0 m) and 0.5-1.5 m deep layer in Jiaozhou Bay. However, the highest growth rate in Shenao Bay appeared under the lowest initial stocking density treatment. In the laboratory, the aquarium experiments (fish and seaweed culture systems) demonstrated that Gracilaria was able to remove inorganic nutrients effectively. The concentration of NH4+-N decreased by 85.53% and 69.45%, and the concentration of PO4-P decreased 65.97% and 26.74% in aquaria with Gracilaria after 23 days and 40 days, respectively. The results indicate that Gracilaria has the potential to remove excess nutrient from coastal areas, and the large-scale cultivation of G. lemaneiformis could be effective to control eutrophication in Chinese coastal waters. (c) 2005 Elsevier B.V. All rights reserved.

Primary culture and characteristic morphologies of medulla terminalis neurons in the eyestalks of Chinese shrimp, Fenneropenaeus chinensis

A method for culturing medulla terminalis (MT) neurons in the eyestalk of Chinese shrimp, Fenneropenaeus chinensis, was first established. The neurons showed immediate outgrowth in the culture medium supplemented with glutamine, glucose and antibiotics. The cells grew for about 2-7 days and then sustained for a week or more. At least six types of neurons were distinguished on the basis of size and form of soma and outgrowth pattern of cells. (C) 2003 Elsevier Science B.V. All rights reserved.

Zingiber officinalis an herbal appetizer in the tiger shrimp Penaeus monodon (Fabricius) larviculture

Penaeus monodon postlarvae were fed with different percentages (0%, 25%, 50%, 75% and 100%) of the herbal appetizer Zingiber officinalis enriched Artemia. After 30 days of culture (i.e. PL-1-30), a very positive result was found in Z. officinalis-enriched Artemia-fed postlarvae. The unenriched Artemia-fed postlarvae consumed 91.0 mg/animal/30 days of feed, whereas the Z. officinalis-enriched Artemia increased their consumption to 127.9 mg/animal/30 days. A similar pattern was noticed in feed absorbed (110.2 mg), dry weight growth (26.7 mg) and feed catabolized (83.2 mg) in Z. officinalis-enriched Artemia because of enzymatic activities. The conversion efficiency of unenriched postlarva was 17.19%, whereas in 100% Z. officinalis-enriched Artemia, the maximum conversion efficiency was 20.85%. The net production efficiency increased significantly (P < 0.05) to 22% from that of the unenriched Artemia-fed postlarvae. The administration of Z. officinalis in all levels produced significantly (P < 0.05) higher weight gain and specific growth rate. The utilization efficiency of feed increased proportionately to the percentages of Z. officinalis. Digestive enzyme activity (amylase, protease and lipase) increased significantly (P < 0.05) in the 50%, 75% and 100% enrichment. Among the different percentages of enrichment, the 100% Z. officinalis-enriched Artemia-fed postlarvae performed better in the overall status.

Comparative growth performances of diploid and triploid Chinese shrimp Fenneropenaeus chinensis (Osbeck, 1765) under different salinities

This study aimed at evaluating the ploidy effects on growth performances of Chinese shrimp (Fenneropenaeus chinensis Osbeck, 1765) reared in different salinities under laboratory conditions. In the acute salinity experiment, there was no difference (P > 0.05) in tolerance observed in triploid and diploid shrimp due to abrupt salinity changes. The lethal salinity for 50% of the individuals in 96 h at 23-25 degrees C was about 2 g L-1 in both triploids and diploids. While for the chronic salinity experiment, statistical analyses confirmed that the differences in growth performances including the specific growth rate (SGR), the feeding rate (FR), feed conversion efficiency (FCE) and intermoult period (IP) between triploid and diploid were related to salinity. Diploid shrimp reared in 20 g L-1 exhibited highest SGR (P < 0.05), while triploids performed well in 20 and 30 g L-1 salinities (P < 0.05). Based on the survival and growth data, the optimal salinity for the culture of diploid F. chinensis should be 20 g L-1 and for triploids it should be between 20 and 30 g L-1.

Molecular cloning, characterization and expression analysis of a putative C-type lectin (Fclectin) gene in Chinese shrimp Fenneropenaeus chinensis

Lectin is regarded as a potential molecule involved in immune recognition and phagocytosis through opsonization in crustacean. Knowledge on lectin at molecular level would help us to understand its regulation mechanism in crustacean immune system. A novel C-type lectin gene (Fclectin) was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 1482 bp with an 861 bp open reading frame, encoding 287 amino acids. The deduced amino acid sequence contains a putative signal peptide of 19 amino acids. It also contains two carbohydrate recognition domains/C-type lectin-like domains (CRD1 and CRD2), which share 78% identity with each other. CRD1 and CRD2 showed 34% and 30% identity with that of mannose-binding lectin from Japanese lamprey (Lethenteron japonicum), respectively. Both CRD1 and CRD2 of Fclectin have I I amino acids residues, which are relatively invariant in animals' C-type lectin CRDs. Five residues at Ca2+ binding site I are conserved in Fclectin. The potential Ca2+/carbohydrate-binding (site 2) motif QPD, E, NP (Gln-Pro-Asp, Glu, Asn-Pro) presented in the two CRDs of Fclectin may support its ability to bind galactose-type sugars. It could be deduced that Fclectin is a member of C-type lectin superfamily. Transcripts of Fclectin were found only in hemocytes by Northern blotting and RNA in situ hybridization. The variation of mRNA transcription level in hemocytes during artificial infection with bacteria and white spot syndrome virus (WSSV) was quantitated by capillary electrophoresis after RT-PCR. An exploration of mRNA expression variation after LPS stimulation was carried out in primarily cultured hemocytes in vitro. Expression profiles of Fclectin gene were greatly modified after bacteria, LPS or WSSV challenge. The above-stated data can provide us clues to understand the probable role of C-type lectin in innate immunity of shrimp and would be helpful to shrimp disease control. (c) 2006 Elsevier Ltd. All rights reserved.

Dynamic changes of total bacteria and Vibrio in an integrated seaweed-abalone culture system

Polyculture of seaweeds alongside fed animal aquaculture is an environmentally friendly means of avoiding eutrophication problem both in land-based and sea-based monoculture systems. Many aspects of such polyculture systems have been described, but little attention has been given to the impact of live seaweeds on the microbiological properties of the water that connects the algae and animals. In this investigation, the Pacific red alga Gracilaria textorii was cultured in a recirculated dual tank system (150 L) with the juvenile abalone Haliotis discus hannai. Dynamic changes of total bacteria (TB) and total Vibrio (TV) in the water of polyculture and monoculture systems were evaluated. Results revealed that (1) level of TB in the polyculture was constantly higher than in the monoculture over a 6.5-day period. While levels of TV in the polyculture was detected to be constantly lower than in the monoculture, (2) integration of G. textorii in the abalone culture changed the Vibrio compositions in the water as judged by the changes of bacteria colony types; (3) application of artificial diet led to dramatic increase of the levels in TB and TV in both systems at 12 h after application in the 24-h test and resulted in selective propagation of Vibrio in the water in the monoculture system; (4) polyculture of G. textorii with juvenile abalone in combination with feeding with live algal diet helped to maintain low levels of TV and the balance of the Vibrio composition; (5) living biomass of G. textorii was effective in preventing propagation of two purified Vibrio strains (V alginolaticus and V logei) in the water. These results provide a general basis of the dynamic changes of levels in TB and TV in a seaweed-abalone polyculture system with or without artificial diet in tanks. (c) 2005 Elsevier B.V All rights reserved.

Evaluation of induced triploid shrimp Penaeus (Fenneropenaeus) chinensis cultured under laboratory conditions

Triploid Penaeus (Fenneropenaeus) chinensis was successfully produced by heat shock. Their metamorphosis, the relationship between body weight and length and difference in appearance between triploids and their diploid siblings under laboratory culture were studied. Hematological studies showed a smaller number of haemocytes, but larger cell volume, in triploids than in diploids. Triploid shrimp did not show higher growth during the immature stage, but exhibited superior growth during the maturation stage. Characteristics of reproductive organs indicated that triploid shrimp may be sterile and sex ratio can be changed through triploidization of shrimp. This paper summarizes the progress made in triploid shrimp research which would be helpful in understanding more about triploids of crustaceans. (c) 2006 Elsevier B.V. All rights reserved.