Population genetic analysis of white shrimp,Litopenaeus setiferus,using microsatellite genetic markers

The white shrimp (Litopenaeus setiferus) is a commercially and recreationally valuable species, yet little is known of its population structure or genetic diversity. White shrimp are distributed along the Atlantic coast of the United States and from the west coast of Florida to the Bay of Campeche, Mexico. In this study, shrimp were collected from North Carolina, South Carolina (four separate collections were taken from 1995 to 1999), Georgia, the Atlantic and Gulf coasts of Florida, Louisiana, Texas and Mexico. DNA was isolated from these individuals, and genetic variation was assessed at six microsatellite loci. These loci were, for the most part, highly polymorphic with an average expected heterozygosity of 0.68. Deviations from Hardy-Weinberg proportions were observed over all samples, but experimental results suggested the presence of null alleles, which confounded a biological interpretation of this result. Pairwise tests of the similarity of allele frequency distributions and distance measure analyses showed broad-scale genetic homogeneity superimposed over occasional indications of random geographical and temporal differentiation. FST and RST estimates over all loci and samples were 0.002 or less and indicated little population structure. Weak but significant genetic differentiation was evident only between pooled western Atlantic and pooled Gulf of Mexico samples. Within the Gulf of Mexico or within the western Atlantic, the large-scale genetic homogeneity observed may be a consequence of genetic mixing resulting from pelagic larvae and adult migrations, while the random local genetic differentiation may be a result of genetic sampling or experimental sampling error. The weak differentiation between shrimp from the Gulf of Mexico and the western Atlantic can be explained by a relatively recent separation of these two populations and/or small amounts of ongoing gene flow.     

Effect of dissolved carbon dioxide on oxygen consumption in the Pacific white shrimp,Litopenaeus vannamei (Boone 1931)

A failure in the aeration system of Litopenaeus vannamei rearing with biofloc technology can decrease the oxygen concentrations rapidly and also increase the carbon dioxide (CO₂) concentrations at the same rate. We report here an evaluation of the effect of CO₂ on the oxygen consumption of L. vannamei. We used a continuous-flow respirometer with water recirculation equipped with a digital fiber-optic oximeter. Eight juveniles of L. vannamei (12.1 ± 1. 4 g) were used in each treatment with one per respiratory chamber (0.6 L). The shrimp were exposed to six concentrations of CO₂ (5, 30, 60, 95, 150, and 300 mgCO₂/L) with an acute exposure time of six hours. Upon treatment with 5–30 mgCO₂/L, we observed a consumption of oxygen of 0.233 ± 0.129 and 0.33 ± 0.072 mgO₂/g/h, respectively. Upon treatment with 60 mgCO₂/L, an increase was observed in the oxygen consumption (0.521 ± 0.098 mgO₂/g/h). Upon treatment with 95, 150, and 300 mgCO₂/L however, the shrimp decreased their oxygen consumption and lost their equilibrium. The CO₂ should therefore be maintained at a maximum of 5 mgCO₂/L during shrimp rearing.     

Characteristics of NADH-dependent lipid peroxidation in sarcoplasmic reticulum of white shrimp,Litopenaeus vannamei,and freshwater prawn,Macrobrachium rosenbergii

The iron-catalyzed NADH-dependent lipid peroxidation system in sarcoplasmic reticulum (SR) of cultured white shrimp, Litopenaeus vannamei and freshwater prawn, Macrobrachium rosenbergii was characterized. Production of thiobarbituric acid reactive substances was used to measure the activity of lipid peroxidation. In both species, the system preferred NADH to NADPH as the reducing agent. Lipid peroxidation activities of SR from both species increased when reaction temperatures increased from 6 to 26 °C. At 66 °C, the reaction was no longer NADH-dependent. Acidic pH amplified the lipid peroxidation activity. Sarcoplasmic reticular lipid peroxidation activity in white shrimp was always greater than in freshwater prawn. Fatty acid composition of SR lipids could be a major factor for this outcome. The proportion of n–3 highly unsaturated fatty acids, such as C20:5 and C22:6, in sarcoplasmic reticular lipids of white shrimp was twice of that in freshwater prawn. The results of this study provide important tools required for anti-oxidative nutrient study at sub-cellular level.     

A major SNP resource for dissection of phenotypic and genetic variation in Pacific white shrimp (Litopenaeus vannamei)

Bioinformatics and re-sequencing approaches were used for the discovery of sequence polymorphisms in Litopenaeus vannamei . A total of 1221 putative single nucleotide polymorphisms (SNPs) were identified in a pool of individuals from various commercial populations. A set of 211 SNPs were selected for further molecular validation and 88% showed variation in 637 samples representing three commercial breeding lines. An association analysis was performed between these markers and several traits of economic importance for shrimp producers including resistance to three major viral diseases. A small number of SNPs showed associations with test weekly gain, grow-out survival and resistance to Taura Syndrome Virus. Very low levels of linkage disequilibrium were revealed between most SNP pairs, with only 11% of SNPs showing an r ²-value above 0.10 with at least one other SNP. Comparison of allele frequencies showed small changes over three generations of the breeding programme in one of the commercial breeding populations. This unique SNP resource has the potential to catalyse future studies of genetic dissection of complex traits, tracing relationships in breeding programmes, and monitoring genetic diversity in commercial and wild populations of L. vannamei .     

Effect of temperature and salinity on the energetics of juvenile gray snapper (Lutjanus griseus): implications for nursery habitat value

Gray snapper (Lutjanus griseus) encounter a wide range of temperatures and salinities in nearshore and estuarine juvenile habitats. The energetic response of juvenile gray snapper to temperature and salinity was measured in laboratory experiments to determine the influence of these physicochemical factors on the potential value of different juvenile nurseries. Maximum consumption and growth rates of juvenile (25-50 mm SL) gray snapper were determined in 12-day trials at 20 temperature/salinity combinations representing conditions in juvenile habitats. Ad libitum feeding level of individual fish was measured daily. Maximum weight specific feeding rate increased significantly with temperature and salinity; however, the effect of salinity was much less than that of temperature. Linear growth rate and specific growth rate both increased with temperature, and salinity did not have a significant effect on either. Gross growth efficiency (K₁, growth×consumption⁻¹*100) increased with temperature and was significantly lower at high salinities, indicating increased energetic costs. The higher K₁ at lower salinities has several implications for juvenile gray snapper in low salinity habitats: (1) they would need less food to achieve the same somatic growth as juveniles in high salinity habitats; (2) they would have higher growth at limited ration levels as compared to high salinity habitats; and (3) they would have less impact on prey populations than higher salinity habitats assuming similar gray snapper densities.     

The effects of exposure to critical thermal maxima on the physiological,metabolic, and immunological responses in adult white shrimp Litopenaeus vannamei (Boone)

Exposure to thermal stress was shown to have a significant effect on the osmotic pressure of the hemolymph, glucose levels, total count of hemocyte (TCH), and proPO activity in adult white shrimp Litopenaeus vannamei. Exposure of the shrimp to CTMax significantly increased the osmotic pressure of the hemolymph relative to the control group. In organisms reaching CTMax, temperature elicited a secondary stress response that included an increase in hemolymph glucose of 31?mg?mL^?1. Metabolites in hemolymph such as cholesterol, acylglycerides, and total protein were not significantly affected by exposure to CTMax. CTMax exposure affected several immunological parameters causing decreases in TCH and proPO activity. We suggested that biomarkers such as osmolality, glucose levels, TCH, and proPO activity could be used as sensitive predictors of exposure to CTMax in white shrimp.     

A response-surface analysis of the relative importance of the temperature,salinity and body weight on the respiratory metabolism of the white shrimp Litopenaeus vannamei (Boone, 1931)

We used a response-surface analysis to determine the importance of different factors affecting the resting routine metabolic rate (QO₂) of the white shrimp Litopenaeus vannamei. The oxygen consumption rates were estimated using a multi-factorial design with 28 combinations of different salinities (15, 20, 25, 30, 35, 40 and 45 psu) and temperature (20, 25, 30 and 35 °C) values. The response-surface analysis produced a quadratic function showing that temperature more profoundly affects the oxygen consumption rate. Response-surface curves were generated to predict the optimal conditions resulting in oxygen consumption to better understand the successful growth of this species.     

An in vitro and in vivo assessment of the potential of Vibrio spp. as probiotics for the Pacific White shrimp,Litopenaeus vannamei

Aims: The objective of the work was to determine whether known strains of nonpathogenic vibrios can act as probiotics for the control of Vibrio infections in the Pacific white shrimp, Litopenaeus vannamei. Methods and Results: Of the ten species tested, only Vibrio alginolyticus (NCIMB 1339) and Vibrio gazogenes (NCIMB 2250) showed antagonistic activity towards a panel of shrimp pathogenic vibrios. In the case of V. alginolyticus, this activity depended on the presence of live bacteria while in V. gazogenes both live and dead bacteria showed anti‐Vibrio activity. Injection of shrimp with either V. alginolyticus or V. gazogenes at 3 × 10⁷ or 3 × 10⁵ total bacteria per shrimp resulted in mortality with higher levels in the case of V. alginolyticus (100% mortality 18 h postinjection of 3 × 10⁷ bacteria). Juvenile shrimp were fed commercial diets top‐coated with either chitin (an immune stimulant) or chitin + V. gazogenes. Both chitin and V. gazogenes caused a significant decline in the number of Vibrio‐like bacteria in the fore and hind gut, and changes were also seen in the hepatosomatic index (a measure of digestive health) and the total number of blood cells in circulation. Analysis of mid/hindgut and faecal samples obtained using terminal restriction fragment length polymorphism showed that the gut microbiota of shrimp has limited bacterial diversity and that after 8 weeks exposure to the experimental diets there were significant changes in the microbial flora of the GI tract of shrimp as a result of the presence of V. gazogenes. Conclusions: Of the vibrios tested, V. gazogenes has potential as a probiotic for the control of bacterial diseases in shrimp. Significance and Impact of the Study: Overall, this study shows the promise of V. gazogenes together with chitin to improve the health and welfare of shrimp under aquaculture conditions.     

Thorn fish Terapon jarbua (Forskål) predation on juvenile white shrimp Penaeus indicus H. Milne Edwards and brown shrimp Metapenaeus monoceros (Fabricius): the effect of turbidity, prey density, substrate type and pneumatophore density

A series of laboratory experiments was conducted at Inhaca Island Marine Biological Station, Mozambique, in order to assess the separate effects of turbidity, prey density, substrate type, pneumatophore density, and the combined effects of turbidity with the latter three, on rate of predation by the thorn fish Terapon jarbua (Forskål, 1775) on white shrimp Penaeus indicus and brown shrimp Metapenaeus monoceros.Significant interactions between turbidity and the other three factors on shrimp predation for both prey species were detected. Regardless of prey density, increasing turbidity decreased predation on P. indicus, but not on M. monoceros, for which increasing densities reduced the protective effect of turbidity. Increasing prey density increased predation on P. indicus in clear water, and increased predation on M. monoceros in low and high, but not in intermediate turbidity or clear water. The presence of a substrate suitable for burying decreased predation on M. monoceros in clear water, but not in the turbidity levels used. In clear water, solely sandy-shell substrate afforded protection to P. indicus, while in turbid water, no substrate offered significant protection and muddy substrate even increased prey vulnerability to fish probably as a result of increased preys' locomotor activity. Raising pneumatophores density seems to lower the protective value of turbidity for both species. In clear water, only low and high structure density provided a deterrent effect on predation on P. indicus; in turbid water, intermediate and higher structure density increased predation. Increasing structural complexity reduced predation on M. monoceros linearly in clear water; but in low turbid water it increased. In high turbid waters, the increase was only significant in intermediate pneumatophore density. High structural complexities impair the pursuing capacity of fish and thus decreased predation rates. The results indicate that the effective provision of shelter of different habitats depends not only on the various environmental parameters analysed, but also on the way they interact and on the behaviour of prey and predator as well.     

Administration of Bacillus subtilis strains in the rearing water enhances the water quality,growth performance,immune response,and resistance against Vibrio harveyi infection in juvenile white shrimp,Litopenaeus vannamei

In this study, vegetative cell suspensions of two Bacillus subtilis strains, L10 and G1 in equal proportions, was administered at two different doses 10⁵ (BM5) and 10⁸ (BM8) CFU ml⁻¹ in the rearing water of shrimp (Litopenaeus vannamei) for eight weeks. Both probiotic groups showed a significant reduction of ammonia, nitrite and nitrate ions under in vitro and in vivo conditions. In comparison to untreated control group, final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR) and digestive enzymatic activity were significantly greater in the BM5 and BM8 groups. Significant differences for survival were recorded in the BM8 group as compared to the control. Eight weeks after the start of experiment, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 80%, whereas cumulative mortality of the shrimp that had been given probiotics was 36.7% with MB8 and 50% with MB5. Subsequently, real-time RT-PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and β-1,3-glucan- binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was only significantly up-regulated in the BM5 group compared to the BM8 and control groups. These results suggest that administration of B. subtilis strains in the rearing water confers beneficial effects for shrimp aquaculture, considering water quality, growth performance, digestive enzymatic activity, immune response and disease resistance.