Chilean Scallop Research Articles

Draft genome sequence of Vibrio lentus VLO8, recovered from the larval culture of the Chilean scallop ( Argopecten purpuratus )

ABSTRACT This announcement reports the genome of Vibrio lentus VLO8 recovered from the larval culture of Chilean scallop. The genomes of strain VLO8 have two contigs with a total length of 5,499,980 bp, an average G + C content of 44.22%, a total number of protein-coding genes of 6,439, and 170 RNAs.

First Report, Characterization and Pathogenicity of Vibrio chagasii Isolated from Diseased Reared Larvae of Chilean Scallop, Argopecten purpuratus (Lamarck, 1819).

Two Vibrio strains (VPAP36 and VPAP40) were isolated from moribund-settled larvae of the Chilean scallop Argopecten purpuratus during vibriosis outbreaks that occurred in two commercial scallop larvae hatcheries located in the Inglesa and Tongoy bays in Northern Chile. The strains were identified as Vibrio chagasii using phenotypic characterization and whole genome sequence analysis. Both strains exhibited the phenotypic properties associated with virulence, gelatin hydrolysis and β-hemolysis, whereas only VPAP36 produced phospholipase and only VPAP40 produced caseinase. The whole genome analysis showed that the strains harbored genes encoding for the virulence factors, the EPS type II secretion system, and Quorum Sensing (auto-inductor 1 and auto-inductor 2), whereas genes encoding a metalloproteinase and a capsular polysaccharide were detected only in the VPAP40 genome. When challenge bioassays using healthy 11-day-old scallop larvae were performed, the V. chagasii VPAP36 and VPAP40 strains exhibited significant (p < 0.05) differences in their larval lethal activity, producing, after 48 h, larval mortalities of 65.51 ± 4.40% and 28.56 ± 5.35%, respectively. Otherwise, the cell-free extracellular products of the VPAP36 and VPAP40 strains produced larval mortalities of 20.86 ± 2.40% and 18.37 ± 2.40%, respectively, after 48 h of exposure. This study reports for the first time the isolation of V. chagasii from the massive larval mortalities of the farmed scallop (Argopecten purpuratus) in Chile, and demonstrates the pathogenic activity of V. chagasii towards the Chilean scallop, the second most important species for Chilean mariculture.

Isolation and Pathogenic Characterization of Vibrio bivalvicida Associated With a Massive Larval Mortality Event in a Commercial Hatchery of Scallop Argopecten purpuratus in Chile.

The VPAP30 strain was isolated as the highly predominant bacteria from an episode of massive larval mortality occurring in a commercial culture of the Chilean scallop Argopecten purpuratus. The main aims of this study were, to characterize and identify the pathogenic strain using biochemical and molecular methods, to demonstrate its pathogenic activity on scallop larvae, to characterize its pathogenic properties and to describe the chronology of the pathology. The pathogenic strain was identified as Vibrio bivalvicida based on its phenotypic properties, the multilocus sequence analysis (MLSA) of eight housekeeping genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA, and topA) and different in silico genome-to-genome comparisons. When triplicate cultures of healthy 10 days old scallop larvae were challenged with 1 × 105 colony forming units (CFU) mL-1 of the VPAP30 strain, percentages of larval survival of 78.9 ± 3.3%, 34.3 ± 4.9%, and 0% were observed at 12, 2,4 and 36 h, respectively, whereas uninfected larval cultures showed survival rates of 97.4 ± 1.2% after of 48 h. Clinical symptoms exhibited by the scallop larvae infected with the VPAP30 strain include the accumulation of bacteria around the scallop larvae, velum disruption and necrosis of digestive gland. The 50% lethal dose (LD50) of VPAP30 strain at 24 and 48 h was 1.3 × 104 and 1.2 × 103 CFU mL-1, respectively. The invasive pathogenic activity of the VPAP30 strain was investigated with staining of the bacterial pathogen with 5-DTAF and analyzing bacterial invasion using epifluorescence, and a complete bacterial dissemination inside the larvae at 24 h post-infection was observed. When scallop larvae were inoculated with cell-free extracellular products (ECPs) of VPAP30, the larval survival rate was 59.5 ± 1.7%, significantly (P < 0.001) lower than the control group (97.4 ± 1.2%) whereas larvae treated with heat-treated ECPs exhibited a survival rate of 61.6 ± 1.8% after 48 h of exposure. V. bivalvicida VPAP30 exhibits high pathogenic activity on scallop larvae, mediated both by bacterial invasion and the production of toxigenic heat-stable compounds. This report constitutes the first isolation of V. bivalvicida out of Europe and extends the host range of this species, having demonstrated its pathogenic activity on the Chilean scallop larvae (A. purpuratus). These results supporting the pathogenic potential of V. bivalvicida to kill the larvae of a broad range of bivalve species reared in hatcheries located in the Atlantic and the Pacific coasts.

Physiological responses of juvenile Chilean scallops (Argopecten purpuratus) to isolated and combined environmental drivers of coastal upwelling

Abstract Coastal biota is exposed to continuous environmental variability as a consequence of natural and anthropogenic processes. Responding to heterogeneous conditions requires the presence of physiological strategies to cope with the environment. Ecosystems influenced by upwelling endure naturally cold, acidic and hypoxic conditions, nevertheless they sustain major fisheries worldwide. This suggests that species inhabiting upwelling habitats possess physiological adaptations to handle high environmental variability. Here, we assessed the impact of the main upwelling drivers (temperature, pH and oxygen) in isolation and combined on eco-physiological responses of Chilean scallop Argopecten purpuratus. A. purpuratus responded to hypoxia by increasing their metabolic performance to maintain growth and calcification. Calcification was only affected by pH and increased under acidic conditions. Further, A. purpuratus juveniles prioritized calcification at the expense of growth under upwelling conditions. Increasing temperature had a significant impact by enhancing the physiological performance of A. purpuratus juveniles independently of oxygen and pH conditions, but this was associated with earlier and higher mortalities. Our results suggest that A. purpuratus is acclimated to short-term colder, acidic and hypoxic conditions, and provide important information of how this species responds to the heterogeneous environment of upwelling, which is significantly relevant in the climatic context of upwelling intensification.

Embryo-larvae and juvenile toxicity of Pb and Cd in Northern Chilean scallop Argopecten purpuratus.

This study aimed to investigate the effects of Cd and Pb on earlier stage of the commercially important scallop (Argopecten purpuratus) in the contamination context of Northern Chile where this specie is farmed, through acute exposure bioassays in embryo-larvae measuring cumulative abnormality (EC50) and juvenile cumulative mortality (LC50) as endpoints, based on environmentally detected concentrations and available toxicological data from similar species. Embryo-larvae exposure indicates 48h EC50 of 1.55mg/L Cd, and 0.044mg/L Pb. On the contrary, 96h LC50 in juvenile scallops was 0.48mg/L Cd and 1.47mg/L Pb. Our results demonstrated differential toxicity between embryo and juvenile scallops that might relate to different primary defense mechanisms or effect in morphological development of individuals in each ontogenetic stage. Compared to similar bivalve metal toxicity tests, this study demonstrated that A. purpuratus embryos are more sensitive to Pb than most other bivalve species. Our results indicate that maximum permitted levels of Pb in marine waters and estuaries (according to Chilean regulation) could pose a risk for scallops' first stage of life (embryo-larvae) development, and needs to be reviewed. Furthermore, Chilean environmental regulations do not have quality standards for marine sediments (currently under discussion), where high levels of metals have been continuously reported.

First Report of Vibrio tubiashii Associated with a Massive Larval Mortality Event in a Commercial Hatchery of Scallop Argopecten purpuratus in Chile.

The VPAP30 strain was isolated as the highly predominant bacteria from an episode of massive larval mortality occurring in a commercial culture of the Chilean scallop Argopecten purpuratus. The main aims of this study were, to characterize and identify the pathogenic strain using biochemical and molecular methods to demonstrate its pathogenic activity on scallop larvae, to characterize its pathogenic properties and to describe the chronology of this pathology. The pathogenic strain was identified as Vibrio tubiashii based on its phenotypic properties and the sequence analysis of its 16S rRNA and housekeeping genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA). When triplicate cultures of healthy 10–day–old scallop larvae were challenged with 1 × 105 colony forming units (CFU) mL-1 of the VPAP30 strain, percentages of larval survival of 78.87 ± 3.33%, 34.32 ± 4.94%, and 0% were observed at 12, 24, and 36 h, respectively; whereas uninfected larval cultures showed survival rates of 97.4 ± 1.24% after of 48 h. Clinical symptoms exhibited by the scallop larvae infected with the VPAP30 strain include the accumulation of bacteria around the scallop larvae, velum disruption and necrosis of digestive gland. The 50% lethal dose (LD50) of VPAP30 strain at 24 and 48 h was 1.3 × 104 and 1.2 × 103 CFU mL-1, respectively. The invasive pathogenic activity of the VPAP30 strain was investigated with staining of the bacterial pathogen with 5-DTAF and analyzing bacterial invasion using epifluorescence, and a complete bacterial dissemination inside the larvae at 24 h post-infection was observed. When scallop larvae were inoculated with cell-free extracellular products (ECPs) of VPAP30, the larval survival rate was 59.5 ± 1.66%, significantly (P < 0.001) lower than the control group (97.4 ± 1.20%) whereas larvae treated with heat-treated ECPs exhibited a survival rate of 61.6 ± 1.84% after 48 h of exposure. This is the first report of the isolation of V. tubiashii from the diseased larvae of the scallop A. purpuratus, occurring in a commercial culture in Chile, and it was demonstrated that the VPAP30 strain exhibits high pathogenic activity on scallop larvae, mediated both by bacterial invasion and the production of toxigenic heat-stable compounds.

Biomineralization changes with food supply confer juvenile scallops (Argopecten purpuratus) resistance to ocean acidification.

Future ocean acidification (OA) will affect physiological traits of marine species, with calcifying species being particularly vulnerable. As OA entails high energy demands, particularly during the rapid juvenile growth phase, food supply may play a key role in the response of marine organisms to OA. We experimentally evaluated the role of food supply in modulating physiological responses and biomineralization processes in juveniles of the Chilean scallop, Argopecten purpuratus, that were exposed to control (pH ~ 8.0) and low pH (pH ~ 7.6) conditions using three food supply treatments (high, intermediate, and low). We found that pH and food levels had additive effects on the physiological response of the juvenile scallops. Metabolic rates, shell growth, net calcification, and ingestion rates increased significantly at low pH conditions, independent of food. These physiological responses increased significantly in organisms exposed to intermediate and high levels of food supply. Hence, food supply seems to play a major role modulating organismal response by providing the energetic means to bolster the physiological response of OA stress. On the contrary, the relative expression of chitin synthase, a functional molecule for biomineralization, increased significantly in scallops exposed to low food supply and low pH, which resulted in a thicker periostracum enriched with chitin polysaccharides. Under reduced food and low pH conditions, the adaptive organismal response was to trade-off growth for the expression of biomineralization molecules and altering of the organic composition of shell periostracum, suggesting that the future performance of these calcifiers will depend on the trajectories of both OA and food supply.Thus, incorporating a suite of traits and multiple stressors in future studies of the adaptive organismal response may provide key insights on OA impacts on marine calcifiers.

Reactive oxygen intermediates promotes the expression of an antimicrobial peptide in the northern scallop Argopecten purpuratus hemocytes in vitro

Reactive oxygen intermediates are metabolites normally produced on aerobic metabolisms and has been linked to processes like oxidative stress and aging. Despite this negative paradigm about ROI, another set of evidence in mammals suggest that these reactive intermediates can act as important second messengers in a variety of cellular signaling processes such as those involved on the immune response. Primary cultures of hemocytes from the Chilean scallop Argopecten purpuratus were developed to study a possible link between the production of reactive oxygen intermediates and the expression of an antimicrobial peptide, big defensin, under a context of immune response. A modified Leibovitz L-15 medium supplemented with 50% of sterile plasma from scallop was essential for hemocyte adhesion an survival. The ability of these primary cultures to develop an immune response was confirmed by phagocytic activity and ROI production essays. The antimicrobial peptide big defensin was up-regulated in response to the β-glucan zymosan and this up regulation could be diminished near to constitutive levels by the treatment of cultured hemocytes with the antioxidant trolox for 3 hours before the immune induction. This results constitutes the first evidence on invertebrates that supports a rol of intracellular oxygen intermediates as promoters of the expression of an antimicrobial effector during an immune response.

Characterization and pathogenicity of Vibrio splendidus strains associated with massive mortalities of commercial hatchery-reared larvae of scallop Argopecten purpuratus (Lamarck, 1819)

Three strains (VPAP16, VPAP18 and VPAP23 strains) were isolated as the most predominant organisms from 3 different episodes of massive mortalities of larval cultures of the Chilean scallop Argopecten purpuratus occurred in different commercial hatcheries located in northern Chile. The main aims of this study were to identify the pathogenic strains and investigate their pathogenic activity. Based on selected phenotypic features and sequence identity of the 16S rRNA gene and the housekeeping gene, RNA polymerase α-chain rpoA, all pathogenic strains were identified as Vibrio splendidus. Healthy 10–day–old scallop larvae cultures exhibited mortality percentages of 69.61±3.35%, 79.78±6.11% and 61.73±3.71% after 48h when were inoculated with 1×106CFU (colony forming units)mL−1 of VPAP16, VPAP18 and VPAP23 strains, respectively, and evidenced that concentrations ⩾104CFUmL−1 would probably be detrimental for the larval culture. The main clinical signs observed in challenged larvae for 24h were bacterial swarms on the margins of the larvae, extension and disruption of the velum, detachment of velum cilia cells and digestive tissue necrosis. Otherwise, challenge assays using pathogenic strains stained with 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein hydrochloride (5-DTAF)evidenced that after 1h stained bacteria were detected in high density in the digestive gland and the margin of the shell. When larval cultures were inoculated with cell-free extracellular products (ECP) of V. splendidus strains, exhibited larval mortalities higher than 70% (VPAP16), 80% (VPAP18) and 50% (VPAP23) after 24h, even when ECP were treated with proteinase K or heat, indicating that extracellular pathogenic activity is mainly mediated by non-proteic thermostable compounds. In this study all Koch’s postulates were fulfilled and it was demonstrated for the first time the pathogenic activity of V. splendidus strains on reared-larvae of scallop A. purpuratus and prompt the necessity to maintain this species at concentrations lower than 104CFUmL−1 to avoid episodes of mass mortalities in scallop hatcheries.

Simulation model of the scallop (Argopecten purpuratus) farming in northern Chile: some applications in the decision making process

Aquaculture farming is a complex system integrating several disciplines, including biology, engineering and economics, all which need to be correctly intertwined to have a profitable and environmentally sustainable activity. During the past recent years, scallop (Argopecten purpuratus) farmers in northern Chile have come to comprehend the hard way that aquaculture producers operate in a complex and dynamic environment where natural and economic factors are in constant change. Thus, to keep a profitable and competitive business in today's world, aquaculture farm managers are in need of relatively easy to use tools for efficient and timely decision making. Harvest size and time, mortality and growth rates, stocking rates, costs and market prices are important variables and parameters to monitor, where decisions with respect to their levels or values have to be made. In this context, non-linear and dynamic quantitative bioeconomic models should become valuable tools, for periodic decision making in the aquaculture business. This paper shows how to emulate Chilean scallop farming using a simulation model that mimics some of the industry's features. The model presented here focuses on a scallop aquaculture center that uses the common technology approach of pearl net and lanterns of the northern region of Chile, and analyses the farming strategies based on harvesting size. Also, these strategies were subject to variations in the parameters in order to identify patterns and asses the sensibility of the model to input values.

Effect of florfenicol and oxytetracycline treatments on the intensive larval culture of the Chilean scallopArgopecten purpuratus(Lamarck, 1819)

The administration of antimicrobials to control bacterial pathologies in Chilean scallop hatcheries is a frequent practice, but their effects on these cultures remained unknown. This study was undertaken to obtain information on the effect of the administration of florfenicol and oxytetracycline on the growth, survival and bacterial content of scallop larvae under farming conditions. Florfenicol-treated cultures exhibited high survival rates (44% after 17 days of culture), whereas cultures not treated or treated with oxytetracycline collapsed after 11 days of culture. Surprisingly, no significant differences in the heterotrophic (Tukey test; P = 0.226) and Vibrio (Tukey test; P = 0.666) concentrations between the oxytetracycline-treated and untreated larval cultures were observed. Otherwise, florfenicol administered directly into rearing tanks produced significantly higher larval growth (Tukey test; P = 0.0001) and survival (Tukey test; P = 0.011) than bath treatment. When 2 and 4 mg L−1 of florfenicol were compared, no significant differences in growth (t-test; P = 0.4596) and survival (Tukey test; P = 0.057) were observed, suggesting that a concentration of 2 mg L−1 is sufficient to ensure larval production. The present results demonstrate the efficacy of florfenicol-based therapy to increase larval survival and growth at commercial scale and prompt the necessity to standardize its use in Chilean scallop hatcheries.

Effect of Increasing Salinity on Weight-Specific Filtration Rate of Juvenile ScallopArgopecten purpuratusReared at Two Temperatures: Is Any Effect Related to Ammonia Buildup?

Larvae of the northern Chilean scallop Argopecten purpuratus (Lamarck, 1819) can be produced in hatcheries in closed aquaculture systems (CAS), and then early juveniles transferred to the sea for further grow-out. A new scallop mass production method from larvae to early juveniles that applies recirculation aquaculture system (RAS) technology has been developed at Universidad Catolica del Norte. RASs might lose water by evaporation, which increases salinity. Water temperature will increase as well if it is not controlled. An experiment was performed to determine the effect of varying salinity and temperature over water quality parameters and scallop physiological processes. Feeding behavior of A. purpuratus was determined under CAS with daily water exchange for hatchery-produced juvenile scallops (mean shell height, 26.5 mm; SD, 1.9) reared at salinities of 34, 38, and 42 g/L, and temperatures at 16°C and 22°C. Weight-specific filtration rates (WFRs) were analyzed at day 12 and again at day 55. Scallops were fed Isochrysis galbana and Chaetoceros calcitrans (ratio, 1:1). Results shows that WFRs analyzed for a 24 h period had a tendency to be lower at 0 h than at 24 h, for both temperatures; WFRs were higher at 22°C than at 16°C. A significant positive regression was also found between final WFR and total ammonia nitrogen (both NH3-N and NH4 -N) levels. Results presented here should be interpreted with caution outside the shell height range of 26.5 ± 1.9 mm. These results are applicable to the management of A. purpuratus under controlled conditions, such as a RAS.

Characterization of culturable bacteria in larval cultures of the Chilean scallop Argopecten purpuratus

The early stages of culture of the Chilean scallop Argopecten purpuratus are affected by the mortality attributed to the presence of pathogenic bacteria. The purpose of this study was to characterize the culturable bacteria present in larval cultures of A. purpuratus in the presence and absence of antibiotic (Tetracycline). Bacterial counts on marine agar from water, algal, and larval samples from the culture units were carried out. These counts were performed every three days during 20 days of incubation. The bacterial strains isolated from count plates were identified by 16S rDNA gene sequence analysis. Additionally, the total bacterial counts from culture water were determined by epifluorescence microscopy. Culturable bacterial counts of water samples ranged from 8 × 103 to 3 × 104 CFU mL–1. These counts represent from 3% to 18% of the total bacterial counts. The culturable bacterial counts in larvae were between 3 × 107 and 3 × 108 CFU g–1 and were on average higher in the absence of antibiotic (P &lt; 0.05). Microalgal bacterial counts ranged from 1 × 104 to 9 × 104 CFU mL–1. Of the 46 strains isolated and identified, 25 (54%) belonged to the Gammaproteobacteria, 11 (24%) to the Bacilli, 5 (11%) to the Actinobacteria, 4 (9%) to the Alphaproteobacteria, and 1 (2%) to the Flavobacteria. The most dominant genera in the water samples were Alteromonas, Brevibacterium, Bacillus, Microbacterium, and Marinobacter, while the most dominant genera in larvae were Pseudoalteromonas, Bacillus, Marinobacterium, Alteromonas, and Vibrio, from a total of 15 different bacterial genera identified. The survival of larvae decreased drastically during the 20 days of culture, showing no differences to antimicrobial treatments. The results indicate that there is a high diversity of culturable bacteria present in the larvae, and most are different from those found in the water and microalgae supplied as food.

Effect of antibiotic treatment on the growth and survival of juvenile northern Chilean scallop,Argopecten purpuratusLamarck (1819), and associated microflora in experimental cultures

Juvenile northern Chilean scallops of 937±55 μm shell height were exposed to five different concentrations of chloramphenicol (CHL) (5, 10, 25, 50 and 100 μg mL−1), plus a control without antibiotics. To determine the effect of CHL on the accompanying microflora, the number of colony-forming units (CFU) that grew on TGE culture medium was counted in the seawater of containers with juveniles, and in containers with microalgae used as food. Both were exposed to the same concentrations of CHL. The growth rates of juveniles treated with CHL and the control without antibiotic showed highly significant differences (P=0.0001). The growth rate was inversely proportional to the CHL concentration. The control sample presented the highest growth rate (84.4±14.3 μm day−1), followed by the sample treated with 5 μg mL−1 (64.2±14.3 μm day−1). The survival in the control and in the treated samples with 5–50 μg mL−1 was rather high, with a mean value of 95%. Only the sample treated with 100 μg mL−1 had a low survival (36.7%). The CFU count was larger in the containers with juveniles, when compared with the ones with food. The CFU count tended to decrease with increasing CHL concentration in the juveniles.

Transcriptomic response of Argopecten purpuratus post-larvae to copper exposure under experimental conditions

Few studies have described the molecular response of mollusk larvae to heavy metal exposure. We investigated the response of Chilean scallop Argopecten purpuratus post-larvae to copper exposure under experimental conditions. Post-larvae were maintained with and without copper stress in tanks containing netlon collectors with biofilms that were formed by the bacterium Halomonas sp. and the diatom Amphora sp., known to increase larval settlement. We focused on the analysis of the differential expression patterns of genes associated with copper response. A suppression subtractive hybridization method was used to identify copper-specific up- and down-regulated genes in the post-larvae following 4 days and 8 days exposure to 2.5 and 10 μg/l Cu +2. This method revealed 145 different sequences corresponding to 10 major physiological functions. The expression of 15 potentially regulated genes was analyzed by real-time PCR in post-larvae at different sampling times during the copper stress. The genes chosen were alpha tubulin, elongation factor 1-alpha ( EF1A), tributylin binding protein type 1 ( TBT), NADH dehydrogenase subunit 2 ( NADH2), cavortin, glutathione peroxidase ( GPx), ferritin, heat shock protein 70 ( HSP70), Pam highwire rpm1 ( Phr1), insulin-like growth factor 1 ( IGF1), cellulase, and ribosomal proteins: L18, L44, S3a and S15. This study contributes to the characterization of potential genetic markers that could be used in future environmental monitoring and to explore new mechanisms of stress tolerance in marine mollusk species, especially in early stages of development.

Non-Destructive Method to Study the Internal Anatomy of the Chilean ScallopArgopecten purpuratus

Scallop aquaculture has a 20-y-old history in Coquimbo, Chile. At the beginning develop and introduce on industrial level the culture techniques was the main goal, but in recent years research to improve the broodstock quality has been introduced. Development of non destructive procedures to determine gonadal ripeness were necessary. The use of magnetic resonance imaging proved to be an interesting tool to view the internal anatomy of the Chilean scallop Argopecten purpuratus (Lamarck 1819) without any harmful side effect. A central groove located in the middle of the adductor muscle, became visible using MRI, and could possibly be related to the haemolymphatic circulation system.

Pathogenicity of a Highly Exopolysaccharide-producing Halomonas Strain Causing Epizootics in Larval Cultures of the Chilean Scallop Argopecten purpuratus (Lamarck, 1819)

Mass mortalities of larval cultures of Chilean scallop Argopecten purpuratus have repeatedly occurred in northern Chile, characterized by larval agglutination and accumulation in the bottom of rearing tanks. The exopolysaccharide slime (EPS) producing CAM2 strain was isolated as the primary organism from moribund larvae in a pathogenic outbreak occurring in a commercial hatchery producing larvae of the Chilean scallop Argopecten purpuratus located in Bahía Inglesa, Chile. The CAM2 strain was characterized biochemically and was identified by polymerase chain reaction amplification of 16S rRNA as Halomonas sp. (Accession number DQ885389.1). Healthy 7-day-old scallop larvae cultures were experimentally infected for a 48-h period with an overnight culture of the CAM2 strain at a final concentration of ca. 10(5) cells per milliliter, and the mortality and vital condition of larvae were determined by optical and scanning electron microscopy (SEM) to describe the chronology of the disease. Pathogenic action of the CAM2 strain was clearly evidenced by SEM analysis, showing a high ability to adhere and detach larvae velum cells by using its "slimy" EPS, producing agglutination, loss of motility, and a posterior sinking of scallop larvae. After 48 h, a dense bacterial slime on the shell surface was observed, producing high percentages of larval agglutination (63.28 +/- 7.87%) and mortality (45.03 +/- 4.32%) that were significantly (P < 0.05) higher than those of the unchallenged control cultures, which exhibited only 3.20 +/- 1.40% dead larvae and no larval agglutination. Furthermore, the CAM2 strain exhibited a high ability to adhere to fiberglass pieces of tanks used for scallop larvae rearing (1.64 x 10(5) cells adhered per square millimeters at 24 h postinoculation), making it very difficult to eradicate it from the culture systems. This is the first report of a pathogenic activity on scallop larvae of Halomonas species, and it prompts the necessity of an appraisal on biofilm-producing bacteria in Chilean scallop hatcheries.

Antibacterial activity of peptide analogs from a synthetic antifungal peptide derived from an immunogenic molecule from Agropecten purpuratus (North Chilean Scallop)

Antibacterial activity of peptide analogs from a synthetic antifungal peptide derived from an immunogenic molecule from Agropecten purpuratus (North Chilean Scallop)

Domestication reduces the capacity to escape from predators

Phenotypic plasticity in response to variations in predatory pressure frequently occurs in wild populations, but it may be more evident and critical in species subjected to high exploitation rates and aquaculture. The Chilean scallop Argopecten purpuratus is becoming a domesticated species and the production of hatchery-reared scallops (closed environment), has implied the development of successive generations of individuals deprived of several stimuli normally present in their natural habitats (e.g. predators). We compared the escape capacities between wild and cultured A. purpuratus and also evaluated the effect of reproductive investment on the escape response capacities. Wild and cultured scallops, at different reproductive stages (maturing, mature and spawned), were stimulated to escape with the predatory sea star Meyenaster gelatinosus. We recorded: (1) the time to reaction, (2) the total number of claps, the duration of the clapping response and the clapping rate until exhaustion, (3) the time they spent closed after exhaustion, and (4) the proportion of claps recovered, the duration of the clapping response and the clapping rate after 20 min of recuperation. We found that wild A. purpuratus (1) reacted earlier when contacted by their natural predator, (2) escaped faster (greater clapping rates), (3) spent less time with their valves closed when exhausted, and (4) most of their escape capacities (i.e. claps number; clapping time; capacity of recuperation) were less affected by the energetic requirements imposed by gonad maturation and/or spawning than in cultured scallops. We considered that all these aspects of the escape response would make wild scallops less vulnerable to predation than cultured scallops, thus decreasing predation risk. Given the reduction of escape performance in cultured scallops, we suggest that this aspect should be considered for the success of culture-based restocking programs.

GENOME SIZE ESTIMATION IN TWO POPULATIONS OF THE NORTHERN CHILEAN SCALLOP, ARGOPECTEN PURPURATUS, USING FLUORESCENCE IMAGE ANALYSIS

Abstract Genome size (the C-value) is known to vary considerably among organisms, but is relatively constant among individuals of the same species. To determine the genome size in the Northern Chilean scallop Argopecten purpuratus, two populations were analyzed by measuring the fluorescence signal in hemocyte nuclei stained with DAPI. The fluorescence intensity was measured during a fluorescence-fading period by image analysis (fluorescence fading method). The area under the curve during a fading period was associated with the genome size, thus we determined the C-value in the Arica population as 1.057 ± 0.057 pg of DNA and 1.139 ± 0.066 pg of DNA for the Tongoy population. The interindividual variation was smaller that 6% for both populations. Variance analysis was performed to detect the effects of organisms and population in the genome size of A. purpuratus. Significant differences were detected between Arica and Tongoy populations (P = 0.0011). The genome size of Tongoy population was larger statistic...