A comparative study of intensive Litopenaeus vannamei culture on four bottom substrates without water change

The effect of four bottom substrates, oyster shell powder (OP), sugarcane bagasse (SB), a mixture of OP and SB (OS) and fresh soil (FS), on the water quality and bacterial and zooplankton density of intensive shrimp (Litopenaeus vannamei) culture tanks without water change and the growth performance of cultured shrimp were compared in this study. At the end of a 110 days culturing trial, the total ammonium-N (TAN) of the water on SB and the nitrite nitrogen (NO₂-N) on OS was significantly lower than that on the other substrates (P<0.05), which coincided with the high density of ammonium- and nitrite-oxidizing bacteria in the water on SB and OS, respectively. The concentration of chlorophyll a (Chl a) increased slowly on OP, SB and OS but remained low on FS. The density of total bacteria on OP, SB and OS was one order of magnitude higher than that on FS, and the density of zooplankton on SB and OS was significantly higher than that on FS or OP (P<0.05). The improved water quality and increased density of bacteria and zooplankton on SB and OS may have had a synergistic effect on shrimp culture, improving its growth performance (high survival rate and yield and low feed conversion rate). SB and OS were more effective for improving the growth performance of intensively cultured L. vannamei without water change than OP and FS. To our knowledge, this study presents the first evidence regarding the effect of different bottom substrates on intensive shrimp culture.     

Partial sequence analysis of mitochondrial COI gene of the chinese shrimp, <Emphasis Type="Italic">fenneropenaeus chinensis</Emphasis>

Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5 and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235 bp (28.0%), 307 bp (36.6%), 138 bp (16.5%) and 158 bp (18.9%), respectively. Furthermore, 556 bp fragment of the sequence was used to discuss the phylogenetic relationship among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp groups: Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus.     

A genetic linkage map of marine shrimp Penaeus (Fenneropenaeus) chinensis based on AFLP, SSR, and RAPD markers

The Chinese shrimp Penaeus (Fenneropaeneus) chinensis is an important species in marine fishery and aquaculture in China. A female Chinese shrimp Penaeus (Fenneropaeneus) chinensis was captured from west coast of the Korean peninsula and mated with a “Yellow Sea No. 1” male to produce the first filial generation (F₁) 100 F₂ full-sib progeny from brother-sister crosses between F₁ families was used for the mapping study. A genetic linkage map of the Chinese shrimp was constructed, based on 354 markers, including 300 amplified fragment length polymorphism (AFLP) markers, 42 microsatellite (SSR) markers, and 12 randomly amplified polymorphism (RAPD) markers. Forty-seven linkage groups (LGs) were identified. The total map length was 4 580.5 cM, with an average spacing of 11.3 cM, covering 75.8% of the estimated genome size. The construction of this genetic linkage map was part of a genetic breeding program. This linkage map will contribute to the discovery of genes and quantitative trait loci (QTLs) in Chinese shrimp.     

Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)

The clotting protein (CP) plays important and diverse roles in crustaceans, such as coagulation and lipid transportation. A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis (named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography. Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca²⁺, suggesting that the clotting reaction is catalyzed by a Ca²⁺-dependent transglutaminase in shrimp hemocytes. The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE. CP exists as disulfide-linked homodimers and oligomers. The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon, Farfantepenaeus paulensis and Litopenaeus vannamei; and similar to that of other decapods. The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry. Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.     

A Study on the contribution of different food sources to shrimp growth in an intensive <Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis> pond

Stable isotope methods can be used to determine the food sources and prey items of aquatic organisms accurately and reliably. This study examined the relative contribution of artificial foods (the formulated feed and Artemia) and natural foods to shrimp growth in an intensive Fenneropenaeus chinensis pond by using carbon and nitrogen stable isotopes. The results showed that the nutrition utilization efficiency of the harvested shrimp was low, only 33.18% of feed nitrogen and 21.73% of feed carbon being converted to shrimp flesh. Our stable isotope results showed that the shrimp obtained nutrition for maximum growth from artificial foods, whose contribution was 93.5%, with the remaining attributed to the natural foods. However, there was 0.94 t harvested shrimp derived from natural foods (the rest of 13.56 t harvested shrimp derived from artificial foods) in 1ha intensive pond with a shrimp production of 14.50 t ha⁻¹. Therefore, unit area shrimp production can be increased by increasing the contribution proportion of natural foods in intensive shrimp farming.     

Effect of bioactive peptides (BPs) on the development of Pacific white shrimp (<Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> Boone, 1931)

The present study was conducted to evaluate the feasibility of replacing fish meal (FM) with bioactive peptides (BPs) in diet of white shrimp (Litopenaeus vannamei). The changes in growth performance, body composition, non-specific immunity, and water quality were examined after the shrimp were fed four diets, in which 0% (control), 33.3%, 66.7% and 100% of FM was replaced by BPs, respectively. The groups were designated as Con, 1/3BPs, 2/3BPs, and 3/3BPs. A total of 720 shrimp with an initial body weight of 1.46 ± 0.78 g were fed the experimental diets for 56 days. The results revealed that: 1) the weight gain rate (WGR) in 1/3BPs, 2/3BPs, and 3/3BPs was significantly higher than that in Con (P < 0.05), while no significant difference was found on survival rate and feed conversion ratio (FCR); 2) the whole-body crude protein (CP) and crude lipids (CL) were significantly different among groups, while there was no significant difference between crude ash and phosphorus contents; 3) the levels of acid phosphatase (ACP), lysozyme (LZM), superoxide dismutase (SOD), phenol oxidase (PO) and bactericidal activity increased significantly with the inclusion of BPs; 4) in terms of water quality, no significant difference was found in pH and dissolved oxygen among diets during the whole experimental period. Moreover, even though nitrite and ammonium levels tended to increase with time, there was no significant difference among groups. The results indicated that BPs is an applicable alternative of protein source, which can substitute FM in the diets of L. vannamei; it is able to effectively promote growth performance and improve immunity. Moreover, BPs in the diets had no negative impact on water quality.     

The brine shrimp (Artemia parthenogenetica) as encapsulation organism for prophylactic chemotherapy of fish and prawn

Brine shrimp (Artemia parthenogenetica) which had ingested three water-insoluble antibacterial drugs i.e. sulfadiazine(SD), oxytetracycline (OTC) and erythromycin estolate (ERY-Es) were fed toTilapia and Mysis IIIof Penaeus orientalis K. The drug contents in the predators were then determined. After administration of drugs toTilapia and Mysis III, through the bio-encapsulation of the brine shrimp, efficacious therapeutical concentration of OTC and ERY-Es (but not SD) in the predators could be reached and maintained for more than 8 hours.     

Species composition and gear characteristics of the <Emphasis Type="Italic">Macrobrachium</Emphasis> fishery of the Cross River Estuary,Nigeria

The Cross River Estuary, Nigeria, is an important shrimping area for artisanal fishermen of the coastal communities. The multi-species Macrobrachium fishery is exploited with three main gears, namely beach seine, push net and trap. Studies on species composition of this fishery recorded thirteen shrimp species, one swimming crab (Callinectes amnicola) and two fish species (Eleotris sp. and Pellonula leonensis). The shrimp species identified included Macrobrachium macrobrachion (83.39% and 55.69% by number and weight, respectively), M. vollenhovenii (9.66% and 37.18%), M. equidens (3.8% and 2.87%), juveniles-sub-adults of Penaeus notialis (1.11% and 1.3%), M. dux, M. felicinum, Palaemonetes africanus, Palaemon maculatus, Palaemon elegans, Desmocaris sp., Leander sp., Nematopalaemon hastatus and Alpheus pontederiae. While the selectivity index for trap was 0.25, beach seine and push net had a lower index of 0.063. The results present the first comprehensive and representative report for the Estuary shrimp fishery and will assist in the management of the biodiversity of this ecosystem.     

Partial Sequence Analysis of Mitochondrial COI Gene of the Chinese Shrimp, Fenneropenaeus Chinensis

Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5 and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235bp (28.0% ),.307bp (36.6%), 138bp (16.5%) and 158bp (18.9%), respectively. Furthermore, 556bp fragment of the sequence was used to discuss the phylogenetic relationship among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp groups: Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus.     

PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp. on intestinal microbial diversity in kuruma shrimp (Marsupenaeus japonicus)

In this study, the intestinal microbiota of kuruma shrimp (Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects of Bacillus spp. on intestinal microbial diversity. Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp. amended feed. PCR and denaturing gradient gel electrophoresis (DGGE) analyses were then performed on DNA extracted directly from the guts. Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S rDNA amplicons, and distinct bands in the gels were sequenced. The results suggested that the gut of kuruma shrimp was dominated by Vibrio sp. and uncultured gamma proteobacterium. Overall, the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity of shrimp.     

The effect of simulated gastrointestinal digestion on shrimp <Emphasis Type="Italic">Penaeus vannamei</Emphasis> allergenicity

Pen a 1 is a major shrimp allergen that induces an allergy. The aim of this study is to assess the effect of simulated gastrointestinal fluids on the allergenicity of Pen a 1. Purified Pen a 1 from shrimp (Penaeus vannamei) was subjected to digestion in SGF (simulated gastric fluid) and SIF (simulated intestinal fluid) for a set time. The allergenicity of digestive Pen a 1 was analyzed by immunoblotting and Ci-ELISA, using pool sera from patients with shrimp specific IgE. The results showed that Pen a 1 exhibited a decrease in allergenicity with increasing digestion time in the SGF and SIF. However, Pen a l exhibited strong resistance to digestive fluids, and all yielded fragments (33 kD, 23 kD, and 14 kD) showed allergic activity. Therefore, anti-digestion may be an important factor for Pen a 1 to induce an allergy.     

Glutamate dehydrogenase and Na+-K+ ATPase expression and growth response of Litopenaeus vannamei to different salinities and dietary protein levels

Improvement in the osmoregulation capacity via nutritional supplies is vitally important in shrimp aquaculture.The effects of dietary protein levels on the osmoregulation capacity of the Pacific white shrimp(L.vannamei) were investigated.This involved an examination of growth performance,glutamate dehydrogenase(GDH) and Na+-K+ ATPase mRNA expression,,and GDH activity in muscles and gills.Three experimental diets were formulated,containing 25%,40%,and 50% dietary protein,and fed to the shrimp at a salinity of 25.After 20 days,no significant difference was observed in weight gain,though GDH and Na+-K+ ATPase gene expression and GDH activity increased with higher dietary protein levels.Subsequently,shrimp fed diets with 25% and 50% dietary protein were transferred into tanks with salinities of 38 and 5,respectively,and sampled at weeks 1 and 2.Shrimp fed with 40% protein at 25 in salinity(optimal conditions) were used as a control.Regardless of the salinities,shrimp fed with 50% dietary protein had significantly higher growth performance than other diets;no significant differences were found in comparison with the control.Shrimp fed with 25% dietary protein and maintained at salinities of 38 and 5 had significantly lower weight gain values after 2 weeks.Ambient salinity change also stimulated the hepatosomatic index,which increased in the first week and then recovered to a relatively normal level,as in the control,after 2 weeks.These findings indicate that in white shrimp,the specific protein nutrient and energy demands related to ambient salinity change are associated with protein metabolism.Increased dietary protein level could improve the osmoregulation capacity of L.vannamei with more energy resources allocated to GDH activity and expression.     

Nitrogen Budget in Recirculating Aquaculture and Water Exchange Systems for Culturing <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

In order to investigate the culture characteristics of two indoor intensive Litopenaeus vannamei farming modes, recirculating aquaculture system (RAS) and water exchange system (WES), this study was carried out to analyze the water quality and nitrogen budget including various forms of nitrogen, microorganism and chlorophyll-a. Nitrogen budget was calculated based on feed input, shrimp harvest, water quality and renewal rate, and collection of bottom mud. Input nitrogen retained in shrimp was 23.58% and 19.10% respectively for WES and RAS, and most of nitrogen waste retained in water and bottom mud. In addition, most of nitrogen in the water of WES was TAN (21.32%) and nitrite (15.30%), while in RAS was nitrate (25.97%), which means that more than 76% of ammonia and nitrite were removed. The effect of microalgae in RAS and WES was negligible. However, bacteria played a great role in the culture system considering the highest cultivable cultivable bacterial populations in RAS and WES were 1.03×10¹⁰ cfu mL^?1 and 2.92×10⁹ cfu mL^?1, respectively. Meanwhile the proportion of bacteria in nitrogen budget was 29.61% and 24.61% in RAS and WES, respectively. RAS and WES could realize shrimp high stocking culture with water consuming rate of 1.25 m³ per kg shrimp and 3.89 m³ per kg shrimp, and power consuming rates of 3.60 kw h per kg shrimp and 2.51 kw h per kg shrimp, respectively. This study revealed the aquatic environment and nitrogen budget of intensive shrimp farming in detail, which provided the scientific basis for improving the industrial shrimp farming.     

Effects of Different Diets on the Dietary Attractability and Selectivity of Chinese Shrimp, Fenneropenaeus chinensis

Attractabilities of different diets and dietary selectivity of Chinese shrimp, Fenneropenaeus chinensis were studied through behavior observation and feeding experiment, respectively. The five diets used in the experiment are: Fish Flesh (FF), Shrimp Flesh (SF), Clam Foot (CF), Polychaete Worm (PW), and Formulated Diet (FD). No significant differences of attractability exist between any two different diets when every two natural diets or all five diets are provided simultaneously. On the other hand, significant differences of attractability exist between FD and every single natural diet when they are provided simultaneously. Results of behavioral observation indicate that natural diets are more attractive than FD. In feeding experiment, Chinese shrimp has distinct selectivity on different diets. It positively selects CF and PW, negatively selects FF and SF, and excludes FD absolutely. The results of the present studies indicate that the dietary selectivity of shrimp was based not only on the attractabilities of the diets, but also on the responses such as growth and food conversion.     

Hematological changes in white spot syndrome virus-infected shrimp, Fenneropenaeus chinensis (Osbeck)

The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.     

Characteristics of the tertiary egg membrane of cuttlefish Sepiella maindroni de Rochebrune

We separated tertiary egg membrane (TGM) from 2- and 25-day-old eggs of cuttlefish Sepiella maindroni de Rochebrune, and revealed its ultrastructure, physical (solubility, barrier property) and biochemical (histology, histochemistry, nutritional components, bacteriostasis) characteristics. The results show that TGM could not be dissolved with natural seawater, alcohol, ether or hydrochloric acid (HCl), but it could be dissolved with 2-chloroethanol, diethylamine, and sodium hydroxide (NaOH). The black TGM was more effective in blocking off mud particulates, microorganisms (Chlorella vulgaris, Vibrio alginolyticus) and lighter than the white TGM. The elasticity of black and white TGMs was 1.8 N and 1.5 N, respectively. There were some ink particulates and rod-shaped bacteria in the black TGM. The nutritional components were different between black and white TGMs: Lipid content was lower and protein content was higher in the black TGM. TGM could also inhibit the growth of Vibrio alginolyticus.     

Chromosomal localization of 5S rDNA in Chinese shrimp (Fenneropenaeus chinensis): a chromosome-specific marker for chromosome identification

Chinese shrimp (Fenneropenaeus chinensis) is an economically important aquaculture species in China. However, cytogenetic and genomic data is limited in the organism partly because the chromosomes are difficult to isolate and analyze. In this study, fluorescence in-situ hybridization (FISH) was used to identify the chromosomes of F. chinensis. The 5S ribosomal RNA gene (rDNA) of F. chinensis was isolated, cloned and then used as a hybridization probe. The results show that the 5S rDNA was located on one pair of homologous chromosomes in F. chinensis. In addition, triploid shrimp were used to evaluate the feasibility of chromosome identification using FISH and to validate the method. It was confirmed that 5S rDNA can be used as a chromosome-specific probe for chromosome identification in F. chinensis. The successful application of FISH in F. chinensis shows that chromosome-specific probes can be developed and this finding will facilitate further research on the chromosomes of penaeid shrimps.     

Effect of co-culture of Chinese shrimp (<Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis>) and sea cucumber (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka) on pond environment

Monoculture of sea cucumber (pond S) and polyculture of shrimp with sea cucumber (pond SS) were established to evaluate the effect of shrimp on the environmental conditions of sea cucumber farming pond. Contributions of sediment organic matter (SOM2) resuspended from benthic sediment and the suspended particulate organic matter (SPOM) deposited from the water column to the precipitated organic matter (SOM1) collected with sediment traps were estimated with carbon stable isotope analysis. The results showed that the levels of SPOM and SOM2 in pond SS significantly decreased in comparison with those in pond S at the end of experiment (P < 0.05), indicating that co-culturing shrimp in sea cucumber farming pond could purify the farming water. Carbon stable isotope analysis showed that the proportion of SOM2 in SOM1 in pond SS (84.97% ± 0.38%) was significantly lower than that in pond S (95.20% ± 0.30%) (P < 0.05), suggesting that the resuspension of organic matter from benthic sediment into overlying water was reduced in polyculture pond. In contrast, the proportion of SPOM in SOM1 in pond SS (15.03% ± 0.38%) was significantly higher than that in pond S (4.80% ± 0.30%) (P < 0.05), indicating that the sedimentation of SPOM from water column was enhanced in pond SS owing to the biodeposition effect of shrimp.