Co-occurrence of white shrimp, Litopenaeus vannamei, mortalities and microcystin toxin in a southeastern USA shrimp facility

Various freshwater and marine algal toxins are known to affect plants, fishes, mammals, and invertebrates. During recent mortality events in Texas white shrimp aquaculture ponds, water and shrimp tissue samples were analyzed for cyanobacterial toxins and found to contain microcystin-LR. Cyanoprokaryota dominated the phytoplankton assemblage in water from the affected pond, particularly Microcystis aeruginosa and Anabaena sp. Water samples from the affected pond also contained high levels of microcystin-LR (45 μg/l), whereas adjacent ponds had a diatom-green algal assemblage and no measurable toxin. Unialgal isolates of M. aeruginosa from the affected pond produced microcystin-LR. Free microcystin-LR concentrations in dead shrimp hepatopancreas determined by HPLC were 55 μg/g total shrimp weight, whereas shrimp hepatopancreas from the adjacent pond without shrimp mortalities had no measurable toxin. Muscle toxin concentration was below 0.1 μg/g.     

AFLP-based genetic linkage map of marine shrimp Penaeus (Fenneropenaeus) chinensis

This paper presents the genetic linkage map of the Chinese shrimp Penaeus (Fenneropenaeus) chinensis constructed with 472 AFLP markers. A hundred F₁ progeny from an intercross between a female from the new variety “Yellow Sea No. 1” and wild caught male used for the mapping study. Two separate maps were constructed for each parent. The female linkage map consisted of 197 marker loci forming 35 linkage groups and spanned a total length of 2191.1 cM, with an average marker space of 13.5 cM. The male map consisted of 194 marker loci mapped to 36 linkage groups and spanned a total length of 1737.3 cM, with an average marker spacing of 11.0 cM. The level of segregation distortion observed in this study was 12.2%. The estimated genome length of P. chinensis was 3150.3 cM for the female and 2549.3 cM for the male, respectively. The observed genome coverage was 69.6% for the female and 68.1% for the male map. The linkage maps constructed in this study provide basic information for further linkage studies on Chinese shrimp, and more importantly, the construction of the maps are part of the work of the genetic breeding programs which will be used for growth discovered in the QTL analysis of P. chinensis.     

Heritability for body weight at harvest size in the Pacific white shrimp, Penaeus (Litopenaeus) vannamei, from a multi-environment experiment using univariate and multivariate animal models

To estimate family BLUP breeding values and the heritability of body weight at harvest size (BW) in the Pacific white shrimp, Penaeus (Litopenaeus) vannamei, an experiment was conduced using information from two farm units of a Mexican hatchery and two shrimp population densities at each location. Data consisted of 12,658 shrimps that were siblings from 48 sires and 77 dams with a nested dam-sire structure. Shrimps were individually weighed at an average age of 130 days post-hatching. BW phenotypic mean (S.D.) was 18.2 (2.4) g, with values ranging from 8.4 to 30.0 g. Data were analyzed using univariate and multivariate models that considered BW within location by density pond environment as a different trait and included or not a common full-sib effect (c). The multivariate animal model included fixed effects of days from hatching and sex. For univariate models that included c effects, BW heritability (S.E.) estimates ranged from 0.24 (0.14) to 0.35 (0.18) across environments (heritability was zero in one environment). For multivariate models (excluding the environment with zero heritability) the heritabilities increased and ranged from 0.37 (0.06) to 0.45 (0.09). Standard errors of heritabilities and c effects were both drastically reduced in the multivariate analysis. Pairwise genetic correlations between environments were from 0.80 (0.08) to 0.86 (0.04). These differences may be indicative of genotype-environment interaction for BW at 130 days post-hatching. Statistical problems found to separate c from additive genetic effects both in univariate models were reduced using multivariate models. Correlation between family raw phenotypic means and family BV means from the multivariate analysis was 0.93 indicating a rather low risk of miss selecting superior families if BLUP solutions were neglected using replicated environment data. It is also concluded that use of incorrect statistical models or unreplicated data may lead to biased or inaccurate estimates of genetic parameters in shrimp breeding programs.     

Serotonin stimulates ovarian maturation and spawning in the black tiger shrimp Penaeus monodon

Serotonin (5-hydroxytryptamine, 5HT) has been reported to induce ovarian maturation and spawning in the crayfish Procambarus clarkii and white Pacific shrimp Litopenaeus vannamei. The aim of this study was to explore the role of exogenous 5HT on the reproductive performance of the black tiger shrimp Penaeus monodon. 5HT solution was injected into domesticated P. monodon broodstock at 50 μg/g body weight and ovarian maturation and spawning were recorded. The presence of 5HT in the ovary and oviduct of P. monodon was also studied by immunohistochemistry and its levels in the ovary by enzyme link immunoabsorbance assay (ELISA). The 5HT-injected P. monodon developed ovarian maturation and spawning rate at the level comparable to that of unilateral eyestalk-ablated shrimp. Hatching rate and the amount of nauplii produced per spawner were also significantly higher in the 5HT-injected shrimp, compared to the eyestalk-ablated shrimp. 5HT-positive reactions were found in the follicular cells of pre-vitellogenic oocytes, in the cytoplasm of early vitellogenic oocytes and on the cell membrane and cytoplasm of late vitellogenic oocytes. 5HT in the ovary was present at 3.53 ± 0.26 ng/mg protein level in previtellogenic stage and increased to 17.03 ± 0.57 ng/mg protein level in the mature stage of the ovary. The results suggest a significant role of 5HT, possibly directly on the ovary and oviduct, on the reproductive function of female P. monodon.     

Chilled storage of white shrimp (Litopenaeus vannamei) spermatophores

Chilled storage of spermatophores from white shrimp (Litopenaeus vannamei) is needed to generate a consistent and reliable supply of spermatozoa for domestication purposes. The objective of this study was to develop a protocol for the chilled storage of white shrimp spermatophores and to evaluate bacterial propagation during such storage. In the first experiment, spermatophores were immersed in four extenders, mineral oil, Ringer's solution, phosphate buffer and 0.85% NaCl, and stored at low temperature (2-4 °C) for 35 days. Characteristics of preserved spermatophores changed the least and viable sperm was highest when spermatophores were stored in mineral oil. Spermatophores preserved with mineral oil appeared morphologically normal. Bacillus circulans, Staphylococcus hominis and S. lugdunensis, S. sciuri, S. xylosus and Micrococcus spp. were identified as the predominant bacteria during chilled storage, and total bacterial counts gradually increased during the experiment. A second experiment investigated the effect of antibiotic on chilled storage. Spermatophores were preserved in only mineral oil or mineral oil with 0.1% penicillin-streptomycin. These were evaluated for changes in external morphology of spermatophores, sperm viability and total bacteria count every week during a 35-day experimental period. Percentages of viable sperm (69.5 ± 3.9%) were significantly higher (P < 0.05) among spermatophores preserved in mineral oil with 0.1% antibiotic compared with those preserved only in mineral oil (57.7 ± 3.4%) over 35 days. The number of total bacteria in the treatment with mineral oil ranged between 28.3 ± 4.8 and 2416.7 ± 299.4 CFU/g, but in mineral oil containing antibiotic bacteria were undetectable. This study suggests that chilled storage of spermatophores is a feasible approach for the management and spawning of white shrimp broodstock.     

Oxytetracycline (OTC) accumulation and elimination in hemolymph, muscle and hepatopancreas of white shrimp Litopenaeus vannamei following an OTC-feed therapeutic treatment

The white shrimp, Litopenaeus vannamei, has become a very important species for the development of shrimp aquaculture in Northwest Mexico. However, viral and bacterial diseases are considered a major threat to the development of this industry. In the present study a trial was conducted to evaluate the tissue distribution, maximum concentration, and elimination of the widely used antibiotic oxytetracycline (OTC) in L. vannamei using indoor tanks under laboratory-controlled conditions. OTC was given to shrimp simulating a therapeutic treatment through medicated feed for 14 days followed by a period of feeding without antibiotic for another 14 days to evaluate the elimination pattern. Samples of hemolymph, muscle, and hepatopancreas were taken from medicated animals every two days for 28 days. All tissues were removed and frozen immediately in liquid nitrogen. OTC levels were analyzed by High Performance Liquid Chromatography (HPLC). Results showed an important OTC increase during consumption of medicated feed in all examined tissues. OTC maximum concentrations were 33.54 ± 11.19, 194.37 ± 16.11, and 18.79 ± 5.87 µg g^− 1 for muscle, hepatopancreas and hemolymph, respectively. Although the highest OTC level was found in the hepatopancreas, it required only two days after the start of dosing to reach this value, whereas the maximum OTC for muscle and hemolymph was detected after eight days of dosing. Ten days after the cessation of medicated feeding, the drug content in the shrimp tail muscle was under the detectable limit for the method (0.01 µg g^− 1 of OTC).     

Morphological, structural, and functional characterization of the haemocytes of the scallop, Argopecten irradians

Argopecten irradians is one of the most important commercial species of Pectinidae family in China. The internal defense system of mollusks consists of circulating haemocytes. In order to characterize the haemocytes of the scallop A. irradians, light and electron microscopical studies were carried out. Four types of haemocytes were recognized: type I small hyalinocytes (2.38 ± 0.08 μm, 30-35%), type II large hyalinocytes (4.41 ± 0.33 μm, 15-20%), type III small granulocytes (4.15 ± 0.26 μm, 20-25%), and type IV large granulocytes (8.26 ± 0.52 μm, 25-30%). Granulocyte types showed smaller N/C ratios than hyalinocytes. The mean haemocyte concentration was about (3.75 ± 0.65) × 10⁷ cells ml^− 1 of haemolymph. Among haemocytes, 44.7% are granular and 55.3% are agranular. These gave a relatively systematic classification scheme for haemocytes of A. irradians. Three types of granules were identified: type I, with high electron-density; type II, with low electron-density; and type III, with a middle level of electron-density, based on TEM studies. Different haemocyte types were not separated with DDGC of Percoll in this study. Both granulocytes and hyalinocytes showed a phagocytic response to the two strains of bacteria, Escherichia coli and RLOs. The phagocytic ability of granulocyte was significantly higher (41-48%) than that of hyalinocyte (9.2-11.2%).     

Effect of high water temperature (33 °C) on the clinical and virological outcome of experimental infections with white spot syndrome virus (WSSV) in specific pathogen-free (SPF) Litopenaeus vannamei

White spot syndrome virus (WSSV) is the most lethal pathogen of cultured shrimp. Previous studies done with undefined WSSV titers showed that high water temperature (32-33 °C) reduced/delayed mortality of WSSV-infected shrimp. This study evaluated the effect of high water temperature on the clinical and virological outcome of a WSSV infection under standardized conditions. Groups of specific pathogen-free Litopenaeus vannamei were challenged either by intramuscular or oral routes with a low (30 SID₅₀) or a high (10,000 SID₅₀) virus titer. Shrimp were kept (i) continuously at 27 °C, (ii) 30 °C or (iii) 33 °C; (iv) maintained at 33 °C before challenge and 27 °C afterwards, or (v) kept at 27 °C before challenge and 33 °C afterwards. Shrimp were maintained at the respective temperatures for 120 h before challenge and 120-144 h post challenge (hpc). Gross signs and mortality were monitored every 12 h until the end of the experiment. Dead and surviving shrimp were screened for WSSV infection (VP28-positive cells) by indirect immunofluorescence (IIF). Shrimp kept continuously at 27 °C or 30 °C, or switched to 27 °C post challenge developed gross signs within 24 hpc, first mortalities at 36-60 hpc and 100% cumulative mortality between 60 and 144 hpc depending on the virus titer. All dead shrimp were WSSV-positive. In contrast, shrimp kept at 33 °C continuously or after WSSV challenge showed no signs of disease and low mortalities (0-30%) regardless of the virus titer. Dead and surviving shrimp were WSSV-negative. Further, early virus replication was studied in two groups of shrimp: one maintained at 27 °C before and after challenge and one switched from 27 °C to 33 °C after challenge with 10,000 SID₅₀. Immunohistochemistry (IHC) analysis showed that WSSV-positive cells were first displayed at 12 hpc in shrimp kept at 27 °C and by 24 hpc the infection became systemic. In contrast, shrimp kept at 33 °C did not display WSSV-positive cells at 12 or 24 hpc. This work confirms previous reports that high water temperature prevents the onset of disease and significantly reduces mortality of WSSV-inoculated shrimp regardless of the route of inoculation or virus titer used. This strategy may have practical applications to control WSSV in tropical shrimp farming countries.     

Detection of V. harveyi in shrimp postlarvae and hatchery tank water by the Most Probable Number technique with PCR

V. harveyi is the cause of serious disease in the shrimp industry in Thailand during cultivation. In this study, the gyrB gene of V. harveyi NICA, isolated from shrimp in Thailand, was sequenced. A pair of specific primers (A2B3) was designed that allowed amplification of a 363 bp gene fragment of V. harveyi. No cross reaction was detected in 17 other Vibrio species tested except for V. carchariae which is a synonym for V. harveyi. The possibility of using A2B3 for confirmation and enumeration of V. harveyi by PCR was demonstrated. Of 40 possible V. harveyi strains isolated from seafood on the basis of their growth on TCBS plates and biochemical reactions, 36 gave a reaction with the specific primers. The primers could detect V. harveyi at a level of as few as 15 cells/ml. The Most Probable Number (MPN) technique was applied to enumerate V. harveyi. We have demonstrated that when PCR was applied directly to the enrichment broth of shrimp artificially inoculated with V. harveyi, the MPN value was no different from the MPN value obtained using the standard technique with selective agar. This technique was employed to enumerate V. harveyi in postlarvae and hatchery tank water. V. harveyi were detected in 18 out of 21 postlarval samples and in 14 out of 21 tank water samples. The numbers of V. harveyi detected in postlarvae and water were 150-1.1 × 10⁸/g postlarvae and 7-4.6 × 10⁴/ml of water samples, respectively. Screening of postlarvae to reduce the high risk of V. harveyi contamination in cultivation ponds is suggested as a measure to prevent the catastrophic losses caused by V. harveyi disease.     

First feeding of Octopus vulgaris Cuvier, 1797 paralarvae using Artemia: Effect of prey size, prey density and feeding frequency

Different assays related to the first feeding of Octopus vulgaris Cuvier, 1797 are compiled in this paper. They include: age at initial feeding age, prey size selection and optimal density, attack timing after feeding, and effect of dose number on the number of captures. Prey capture and ingestion processes were also analysed. Food supplied was cultured Artemia sp. Each assay lasted 15 min.Although paralarvae already start to feed on the hatching day (day 0), it is during day 2 when a greater number of attacks is recorded (81.7 ± 14.7% paralarvae attack). They mainly prefer (significance level α = 0.05) large Artemia, 1.4 ± 0.4 mm (77.0 ± 5.6% of the total attacks) than small Artemia, 0.8 ± 0.1 mm (23.0 ± 5.6%). There is also a slight predilection for the lowest Artemia concentration (33.3 ±12.6% paralarvae attack in a 0.1 Artemia ml^− 1 density, opposite 16.7 ± 7.6 and 18.3 ± 7.6% in densities of 0.5 and 1 Artemia ml^− 1 respectively). The greatest predatory activity is recorded during the first 5 min after food is supplied (72.2 ± 25.5%). An increase in the predatory activity was also observed when food was distributed in several doses instead of a single dose (75.0 ± 10.0% and 46.7 ± 17.6% respectively). It was proved for the first time that paralarvae completely ingest their preys (including their exoskeletons), in this case Artemia. Time needed for their total ingestion ranges between 4 and 10 min.     

Growth, immune function, and disease and stress resistance of juvenile Nile tilapia (Oreochromis niloticus) fed graded levels of bovine lactoferrin

Juvenile Nile tilapia (Oreochromis niloticus) were fed nutritionally complete, practical basal diets supplemented with bovine lactoferrin (Lf) at 0, 200, 400, 800, or 1600 mg/kg diet to apparent satiation twice daily for 8 weeks. After the feeding trial, the effect of dietary Lf on growth performance, immune function, and resistance to Streptococcus iniae challenge and low-water stress was determined. Dietary Lf did not affect growth performance (weight gain, feed intake, feed efficiency ratio, or survival) or haematological parameters (haemoglobin, white and red blood cell counts, or haematocrit) (P > 0.05). Crowding stress produced significant increases in plasma cortisol, glucose, lactate, and osmolality from baseline values (P ≤ 0.001), but dietary Lf did not affect plasma glucose, osmolality, or cortisol concentrations (P > 0.05). The level of Lf in diet had a significant impact on survival following S. iniae challenge with fish fed the 800 mg/kg Lf diet having significantly higher survival than control fish (P ≤ 0.05). There was not a corresponding increase in activity of non-specific or specific immune parameters (plasma lysozyme and spontaneous haemolytic complement activities or agglutination antibody titer against S. iniae) with addition of Lf to diets (P > 0.05), but plasma iron decreased and total iron binding capacity (TIBC) increased significantly with increasing concentration of Lf in diet (P ≤ 0.05). The ability of Lf to sequester iron, an essential nutrient required for the growth of bacteria, is regarded as one of its key antibacterial properties. The increased survival with increasing dietary concentration of Lf seemed to correspond with a decrease in plasma iron concentration and not enhancement of non-specific or specific immune functions.     

A stochastic approach for analysis of the influence of white spot disease, zootechnical parameters, water quality, and management factors on the variability of production of shrimp Litopenaeus vannamei cultivated under intensive commercial conditions

We investigated the variability of shrimp Litopenaeus vannamei production by incorporating stochastic elements into deterministic stock models and determined the contribution that white spot disease, zootechnical parameters, water quality, and alternative management strategies have on variability. The model was calibrated for intensive shrimp cultivation in the State of Nayarit, Mexico. Mean annual production increased as a consequence of improved management from 8000 kg ha⁻¹ to 22,000 kg ha⁻¹ when cultivation was not affected by the disease and from 3200 kg ha⁻¹ to 10,400 kg ha⁻¹ when the disease affected production. When simultaneously considering both cases, mean annual production increased from 6300 kg ha⁻¹ to 16,800 kg ha⁻¹. White spot disease was a major factor determining variability of production. Shrimp production was particularly sensitive to levels of dissolved oxygen when management was inadequate, while final weight and mortality rate of shrimp were more sensitive when management improved. Water temperature and salinity had intermediate importance, and mortality caused by the disease and the time when mortality occurred had intermediate or low relevance. Improving management increased shrimp production and diminished variability. The duration of cultivation and stocking density were the most important management variables controlling variability of production when cultivation was affected by the white spot disease. When the disease was not present, pond size and duration of cultivation were the main factors affecting production. Starting time of aeration had relatively lower importance in determining variability, while the stochastic values of dissolved oxygen, in contrast, became most important. These results call for studies on improving aeration management to reduce variability of dissolved oxygen in ponds.     

The mariculture potential of the Indonesian reef-dwelling sponge Callyspongia (Euplacella) biru: Growth, survival and bioactive compounds

In the present study, the Indonesian reef-dwelling sponge Callyspongia (Euplacella) biru de Voogd [de Voogd, N.J., 2004. Callyspongia (Euplacella) biru spec. nov. (Porifera: Demospongiae: Haplosclerida) from Indonesia. Zool. Meded. 78, 477-483.] (Demospongiae, Callyspongiidae) was cultured for a period of 6 months at three sites in the Spermonde Archipelago, Indonesia. A total of 250 sponge cuttings of two size classes (30 and 40 mm in length) were threaded on either polyethylene or very fine nylon rope and attached to horizontal mooring systems. All sponge cuttings were photographed at regular time intervals. Survival rates were high (82%-100%) among all treatments used. Growth rates differed significantly among treatments, but did not differ significantly per location, farming method, explant size and explant position obtained from the parent sponge. The concentration of the bioactive compound amphitoxin in the cultured explants was not significantly lower than in the natural population, but the concentration did vary significantly between explants used in the different treatments, and also between the different fragments of the natural population. The high growth potential and high survival rate suggest that this sponge species is a promising candidate for further mariculture development.     

Effects of salinity and pH on ion-transport enzyme activities, survival and growth of Litopenaeus vannamei postlarvae

Effects of the salinity and pH on ion-transport enzyme activities, survival and growth of Litopenaeus vannamei postlarvae were investigated. Shrimp were transferred from salinity 31‰ and pH 8.1 to different salinity levels of 22, 25, 28, 31 (control), and to different pH levels of 7.1, 7.6, 8.1 (control), 8.6 and 9.1. The results showed ion-transport enzyme activities and weight gains of postlarvae were significantly affected by salinity and pH variation, which had no obvious effect on survival rate. The changing salinity affected the activities of total ATPase and Na⁺-K⁺-ATPase notably (F > F_0.05), meanwhile, non-significantly to the activities of V-ATPase, HCO₃⁻-ATPase. Within 48 h of salinity changing, the activities of ATPase, Na⁺-K⁺-ATPase in each treatment group gradually increased with the sampling time and reached their climax at 48 h, and then stabilized, showing negative correlation with salinity. The changing of pH affected greatly the activities of ATPase, Na⁺-K⁺-ATPase, V-ATPase and HCO₃⁻-ATPase (F > F_0.05), the activities of ATPase, Na⁺-K⁺-ATPase in each treatment group (pH = 7.1, 7.6, 8.6, 9.1) showed peak change within 72 h and stabilized afterwards, and the Na⁺-K⁺-ATPase activities came back to the level of the control group; Meanwhile the changing extent of V-ATPase and HCO₃⁻-ATPase activity corresponded with the grads of pH, and these ATPase activities showed negative correlation with pH changing, the activities of V-ATPase, HCO₃⁻-ATPase in postlarvae of each treatment group came to stable level after 24 h. The experiment also indicated the strength order of these ion-transport enzyme activities were as follows: Na⁺-K⁺-ATPase > V-ATPase > HCO₃⁻-ATPase. Na⁺-K⁺-ATPase was the chief undertaker of osmoregulation under the salinity effects, while V-ATPase and HCO₃⁻-ATPase were the chief osmoregulation undertakers under pH changing. In different salinity environment, the contributions of Na⁺-K⁺-ATPase, V-ATPase and HCO₃⁻-ATPase of L. vannamei postlarvae approximately accounted for 62.0-78.0%, 15.9-29.0% and 2.03-4.12% of ATPase activities in total, respectively. Meanwhile, in different pH medium, the contributions of these ATPases approximately accounted for 50.7-67.4%, 21.3-31.8% and 2.15-7.90% of total ATPase activities, respectively. Weight gain of shrimp transferred to salinity 31 (control) and 28‰ was significantly higher than that of shrimp reared at 25 and 22‰, and weight gain of shrimp transferred to pH 8.1 (control) and 8.6 was significantly higher that that of shrimp transferred to pH 7.1, 7.6 and 9.1. It was suggested that during the process of desalting and culturing of postlarvae, the salinity changing should not exceed 3 and pH variety not more than 0.5.     

Ammonia toxicity and its effect on the growth of the South African abalone Haliotis midae Linnaeus

The current study investigated acute toxicity to ammonia of the South African abalone, Haliotis midae, from three size classes relevant to mariculture operations, and the chronic impact of sub-lethal ammonia levels on growth of juvenile abalone.Results showed that tolerance to ammonia (at pH 7.8 and T_a = 15 °C) increases with body size (i.e. age) as indicated by 36 h LC₅₀ values: juvenile abalone (1-2.5 cm shell length) had the lowest LC₅₀ of 9.8 μg l^− 1 FAN, whereas LC₅₀ was 12.9 μg l^− 1 FAN in “cocktail”-size abalone (5-8 cm shell length). The highest LC₅₀ of 16.4 μg l^− 1 FAN was observed in “brood stock”-size animals (10-15 cm). When “cocktail”-size abalone were allowed to acclimatize to sub-lethal ammonia levels for 48 h, their ammonia tolerance increased compared with non-acclimatized abalone of the same size: LC₅₀ was 2.0 μg l^− 1 FAN higher at 14.8 μg l^− 1 FAN.Growth of juvenile abalone (1-2.5 cm shell length) during chronic exposure to sub-lethal FAN levels is inhibited: specific growth rate (SGR) is significantly reduced by 58.7% to 0.10 ± 0.03% d^− 1 (weight) compared with 0.24 ± 0.06% d^− 1 of abalone of a control group (no ammonia).The results demonstrate the negative effects of ammonia not only on survival but also on growth of farmed abalone, both impair profitability of the farming operation. The information from the present study will be helpful in determining water quality requirements in South African abalone farms.     

Nitrogen and phosphorous budgets during a farming cycle of the Manila clam Ruditapes philippinarum: An in situ experiment

In the Sacca di Goro lagoon a farming cycle of the Manila clam (Ruditapes philippinarum) was simulated seeding young molluscs in an unexploited sandy spot. The experimental area (2100 m²) consisted of three sectors: a control (C), almost devoid of clams (∼ 1600 m^− 2, ∼ 30 ind m^− 2), a low (L) density area (400 m², ∼ 300 ind m^− 2) and a high (H) density zone (∼ 110 m^− 2, ∼ 800 ind m^− 2). Water chemistry, external freshwater nutrient loads, molluscs filtration rates, biomass, elemental composition and nutrient recycling were analysed.Clam filtration rates and light and dark fluxes of nutrients were measured with intact core incubations. Three replicate cores (i.d. 20 cm) were collected from C, L and H in April, one month after the seeding, June, August and October 2003. External loads were calculated multiplying dissolved and particulate nutrients concentration by freshwater flow from the main lagoon tributaries. Direct excretion, filtration activity of clams and particulate matter deposition resulted in significantly higher ammonium (NH₄⁺) and soluble reactive phosphorus (SRP) effluxes to the water column at L and H. For the entire farming cycle, particulate nitrogen (PN) uptake by clams from the water column was 1.7 (C), 9.1 (L) and 16.3 (H) mol m^− 2, whilst total dissolved nitrogen (TDN) fluxes were − 0.3 (C), 1.6 (L) and 6.9 (H) mol m^− 2. Particulate phosphorus (PP) uptake from the water column was 0.1 (C), 0.6 (L) and 1.0 (H) mol m^− 2, whilst total dissolved phosphorus (TDP) efflux was 0.2 (C), 0.5 (L) and 0.8 (H) mol m^− 2. At the end of the farming cycle, harvested N as mollusc flesh was negligible for C, 0.4 mol m^− 2 for L and 1.8 mol m^− 2 for H. Harvested P as mollusc flesh was negligible for C, 0.02 mol m^− 2 for L and 0.04 mol m^− 2 for H. Farmed areas seem to have a great potential for fast coupling between sedimentation (filter feeder mediated biodeposition) and benthic recycling. At the lagoon level, mollusc farming probably attenuates the export of particulate matter to the open sea. Our results show that a minor fraction of biodeposited N (∼ 6%) and P (∼ 3%) was exported as a commercial product at the end of the farming cycle, whilst a larger fraction was incorporated in the sediments or recycled as dissolved inorganic or organic forms.     

Analysis of viral load between different tissues and rate of progression of white spot syndrome virus (WSSV) in Penaeus monodon

At present the most common and most devastating disease of shrimp is caused by the white spot syndrome virus (WSSV), which has spread throughout the world mainly by different species of crustaceans carrying the virus. After experimental injection of Penaeus monodon with a known copy number of WSSV in the abdominal muscle, the rate of viral progression in different tissues at 12, 24, 36 and 48 hpi (hours post infection) was assessed using quantitative real‐time PCR. At 12 hpi the viral load was highest in haemocytes followed by pleopod, muscle and gills whereas at 48 hpi, the gills, the main target of WSSV, showed the highest viral load followed by pleopod, muscle and haemocytes. Viral copy number in the haemocytes was the lowest beyond 12 hpi indicating a remarkable reduction in the rate of viral replication in haemocytes compared with other tissues. The viral load in haemocytes, though increased again beyond 36 hpi, never surpassed the load in the other tissues. The real‐time PCR assay with its high sensitivity and wide dynamic range make it ideal for detecting low‐level WSSV infections that can occur in apparently healthy P. monodon.     

Studies on the rotifer (Brachionus urceus Linnaeus, 1758) as a vector in white spot syndrome virus (WSSV) transmission

The rotifer Brachionus urceus (Linnaeus, 1758) was experimentally infected with the white spot syndrome virus (WSSV) by the virus-phytoplankton adhesion route in order to assess the possibility of rotifer acting as a vector of WSSV to infect the shrimp Fenneropenaeus chinensis (Osbeck, 1765) larvae at zoea stage III. The nested-PCR test revealed WSSV-positive results in the rotifers exposed to WSSV by the virus-phytoplankton adhesion route. Among 10 replicates in the infection treatment, 40% of F. chinensis larvae became WSSV-positive when fed with WSSV-positive rotifers, whereas all were WSSV-negative for F. chinensis when fed with WSSV-free rotifers. Though the mortality of shrimp larvae in the infection treatment (39.47 ± 15.44%) was higher than that in the control treatment (34.67 ± 15.11%), there was no significant difference in the mortality between them (P > 0.05). These results indicated that the rotifer could serve as a vector in WSSV transmission when ingested.     

凡纳滨对虾AP-1基因的克隆和表达特征分析

为了探讨凡纳滨对虾转录因子AP-1在病毒引发的免疫应答过程中的潜在作用,实验根据前期的转录组和表达谱结果提示信息,首次克隆了凡纳滨对虾AP-1基因(LvAP-1,GenBank注册号:KF999956),利用在线软件进行了生物信息学分析,运用半定量的方法进行了组织表达分析,并利用实时荧光定量PCR(qPCR)技术分析了该基因在白斑杆状病毒(WSSV)侵染过程中的表达变化特征。结果显示,AP-1基因ORF区全长882 bp,编码293个氨基酸。预测分析显示该基因编码的蛋白质含有1个Jun结构域和1个高度保守的亮氨酸拉链结构域(bZIP),其中Jun结构域在非脊椎动物中保守性不高。组织表达分析表明,该基因在凡纳滨对虾各组织中广泛表达,其中在血细胞中表达量最高。在WSSV感染早期(0.5 hpi),该基因表达没有显著改变,感染后5 h(5 hpi),AP-1基因开始显著上调表达,在人工感染后24 h,该基因的表达量达到最高(P0.01)。研究表明,该基因在一定程度上参与了凡纳滨对虾体内由WSSV引发的先天免疫应答过程,为进一步研究LvAP-1在对虾应答病毒侵染过程中的功能和作用机制奠定了基础。     

Oxytetracycline (OTC) uptake following bath treatment in gilthead sea bream (Sparus aurata)

The uptake and elimination profile of oxytetracycline (OTC) following a prolong bath treatment in gilthhead sea bream (Sparus aurata) were investigated in this study. The bath experiment was carried out using a OTC concentration of 50 μg/ml for 24 h at 17-18 °C water temperature. Plasma and muscle fish samples were analysed at 1, 3, 6 and 24 h during and at 1, 2, 3, 4 and 6 d following the bath. Detectable OTC levels were revealed only at the end of bath treatment (24 h) in examined tissues of gilthead sea bream, where drug concentration was measured to be as low as 0.096 and 0.047 μg/g or ml in muscle plus skin and plasma, respectively. The findings of the present study indicate that OTC bath treatment under this dosage schedule is unlikely to confront systemic bacterial infections.