Effects of formalin on total haemocytes count and histopathological changes in the shrimp Litopenaeus schmitti (Pérez‐Farfante & Kensley 1997)

Formalin is used in shrimp industry to eliminate ectoparasites and as a disinfectant. However, the effect of formalin on shrimp's defense mechanism is unknown. Several biomarkers were used to assess the immune response of juveniles of the shrimp Litopenaeus schmitti exposed to different concentrations of formalin (10, 25 and 50 mg L^?1) during 24 and 48 h. Formalin concentrations of 25 and 50 mg L^?1 produced a significant reduction in the total haemocyte count at both 24 and 48 h. Peroxidase and phenoloxidase activity did not show variations. The total protein concentration in haemolymph of shrimps was not significantly different between experimental groups. Histological analysis showed hyperplasia in the gills of animals exposed at 10 mg L^?1 for 24 h; however, the exposure at 25 and 50 mg L^?1 caused necrosis in gills, hepathopancreas and the antennal gland.     

Effects of low salinity exposure on immunological,physiological and growth performance in Litopenaeus vannamei

Growth, immunological and physiological parameters of white shrimp Litopenaeus vannamei reared at different salinity levels (1, 10, 15, 25 and 35 g/L) at stocking density of 214 shrimp/m³ were examined at 1, 30 and 63 days. Results showed that the total haemocyte count (THC) of shrimp decreased with time at all salinity levels, indicating a potential reduction in the resistance of shrimp against pathogens, since a low value of THC indicates a perturbation of the immune system. Glucose and protein values observed in the haemolymph throughout the study indicate that shrimp adapted well to low salinities (1, 10 and 15 g/L). Although of those shrimp reared at 10 g/L only 83.3% survived, at this salinity, shrimp depicted a higher glucose concentration in haemolymph at the beginning and end of the study.     

Comparison of immune responses and antioxidant status of different generations of growth‐selected Portunus trituberculatus families

The purpose of this study was to investigate the effects of inbreeding on the immune responses and antioxidant status of Portunus trituberculatus juveniles. Results showed that inbreeding affected the total haemocyte count, and phagocytic, pro‐phenoloxidase (propo), phenoloxidase and antibacterial activities decreased after the seventh generation. Antioxidant status showed a similar pattern: total antioxidant capacity, superoxide dismutase (SOD) activity and GSH/GSSG in the cell‐free haemolymph and hepatopancreas decreased, while catalase activity in the cell‐free haemolymph increased (P < 0.05). There were no significant differences in α₂‐macroglobulin and bacteriolytic activities in the cell‐free haemolymph and glutathione peroxidase activity in the cell‐free haemolymph and hepatopancreas among nine inbreeding generations. Gene expression levels of proPO and crustin in haemocytes and SOD in haemocytes and hepatopancreas also decreased significantly as the inbreeding generations increased. The results suggest that a high level of inbreeding could severely reduce the physiological health of P. trituberculatus. Our obtained data would be particularly useful for P. trituberculatus breeding programmes.     

Effect of hypo- and hyper-saline conditions on osmolarity and fatty acid composition of juvenile shrimp Litopenaeus vannamei (Boone, 1931) fed low- and high-HUFA diets

Litopenaeus vannamei (Boone) grown in ponds are exposed to salinities of less than 5 g L^?1 during inland shrimp culture or to more than 40 g L^?1 from evaporation and reduced water exchange in dry, hot climates. However, dietary requirements for shrimp grown in low or high salinities are not well defined, particularly for fatty acids. Feeding shrimp postlarvae with highly unsaturated fatty acids (HUFA) enhances tolerance to acute exposure to low salinity, as a result of better nutritional status, or/and specific effects of HUFA on membrane function and osmoregulation mechanisms. This study analysed the effect of HUFA supplementation (3% vs. 34%) on L. vannamei juveniles reared for 21 days at low (5 g L^?1), medium (30 g L^?1) and high salinities (50 g L^?1). Juveniles grown at 5 g L^?1 had lower survival compared with controls (30 g L^?1) or shrimp grown at 50 g L^?1, but no significant effect on survival was observed as a result of HUFA enrichment. In contrast, growth was significantly lower for shrimp grown at 50 g L^?1, but this effect was compensated by the HUFA‐enriched diet. Osmotic pressure in haemolymph was affected by salinity, but not by HUFA enrichment. Shrimp fed HUFA‐enriched diets had significantly higher levels of eicosapentaenoic acid and docosahexaenoic acid in hepatopancreas and gills. These results demonstrate that growth at high salinities is enhanced with diets containing high HUFA levels, but that HUFA‐enriched diets have no effect on shrimp reared at low salinities.     

Effects of dietary β‐glucan and glycyrrhizin on non‐specific immunity and disease resistance of white shrimp,Litopenaeus vannamei (Boone) challenged with Vibrio alginolyticus

The white shrimp Litopenaeus vannamei, fed immunostimulant‐free, 0.2%β‐glucan and 0.06% glycyrrhizin diets for 18 days, respectively, were challenged with Vibrio alginolyticus at 6.4 × 10⁴ CFU shrimp^?1. The total haemocyte count (THC), phenoloxidase (PO) activity, respiratory burst (RB) and superoxide dismutase (SOD) activity changes for a 120‐h period were investigated, and shrimp mortality was also recorded. The results showed that PO activity, RB and SOD activity were significantly higher in shrimp fed the two immunostimulant diets after 18 days than those in shrimp fed immunostimulant‐free diets. The THC and SOD activity decreased significantly from 0 to 24 h post challenge, and then reverted to normal levels at 96 and 72 h respectively. The values for PO activity and RB increased from 0 to 48 h post challenge. Compared with those fed the control diets, shrimp fed immunostimulants had significantly higher PO activity and RB values at 120 h post challenge. Mortalities after challenge with V. alginolyticus were significantly lower in shrimp fed with β‐glucan or glycyrrhizin than in those fed with a diet without immunostimulants. It was concluded that dietary β‐glucan and glycyrrhizin increased the shrimp immunity. Furthermore, β‐glucan caused an increase in some immune parameters 12 h earlier than glycyrrhizin after V. alginolyticus challenge.     

Influence of temperature and salinity on the yellowleg shrimp, Penaeus californieinsis Holmes, prophenoloxidase system

The effects of salinity and temperature on plasma protein concentration and total haemocytic prophenoloxidase (proPO) were determined. Groups of 10 juvenile yellowleg shrimp, Penaeus californiensis Holmes, were acclimated for 20 days at different salinities (28%o, 32%o, 36%o, 40%o and 44%o at 25 °C) or temperatures (18, 22, 25, 28 and 32 °C at 36%o). While total protein levels were not affected, the quantities of proPO increased as salinity was elevated. Temperature affected both haemolymph parameters, showing a significant decrease in proPO at 32 °C, and an increase of protein at 28 and 32 °C. These results may help to explain the elevated disease susceptibility of shrimp at higher salinities and/or temperatures.     

Lambari fish Deuterodon iguape as an alternative to live bait for estuarine recreational fishing

Fishing tourism is growing steadily in Brazil. Artisanal fishers meeting the demand for recreational fishing are capturing live juvenile shrimp in the estuaries and selling them for live bait, preventing the recruitment of a larger contingent into the ocean. This study compares the efficiency of using lambari, Deuterodon iguape Eigenmann, as live bait with the shrimp, Penaeus schmitti Burkenroad, while fishing for common snook (Centropomus spp.), as well as to evaluate the swimming behaviour of D. iguape in different salinities. The results showed that there was no significant difference between the live baits used (P. schmitti vs D. iguape). The month, the period of the day and salinity presented differences. The size of the species did not depend on the type of bait used. There was no statistical difference in swimming behaviour, but there was a trend of increased speed and displacement in the group exposed to a salinity of 10 g/L. In conclusion, D. iguape appears as effective as P. schmitti and can be successfully used as live bait; and from an ecological point of view, it reduces the impact of fishing on juvenile shrimp.     

A chimeric recombinant crustacean hyperglycemic hormone from Litopenaeus schmitti (Burkenroad) obtained as C‐terminus fusion protein boost hemolymph glucose concentration in Litopenaeus vannamei (Boone)

To date, no hormonal treatments are available for control of shrimp reproduction and only eyestalk ablation is of practical use. The crustacean hyperglycemic hormone (CHH) is the most abundant neuropeptide of the eyestalk CHH family. It plays an important role in the regulation of hemolymph glucose levels, as its principal function, but it is also implicated in additional physiological processes such as moulting and reproduction. In the present study, the cDNA encoding Litopenaeus schmitti (Burkenroad) mature CHH was cloned into the Escherichia coli pTYB2 expression vector. Using this strategy we have obtained, for the first time, the recombinant CHH from L. schmitti with its C‐terminus fused to an intein tag. The expected fused protein of about 63 KDa was expressed in E. coli forming inclusion bodies. It was purified in a soluble form by electroelution following molecular size fractionation in sodium dodecil sulphate polyacrylamide gel electrophoresis. The ability of the chimeric CHH protein to elevate glucose levels in the hemolymph of the eyestalk‐ablated Litopenaeus vannamei (Boone) shrimps indicates that its biological activity as hyperglycemic protein is preserved. The results provide an alternative tool to obtain soluble recombinant proteins from the CHH family of neuropeptides to get a better understanding of shrimp endocrinology.     

Effect of promoted natural feed on the production,nutritional, and immunological parameters of Litopenaeus vannamei (Boone, 1931) semi‐intensively farmed

The effect of promoted biota on the production parameters, water quality, nutritional and immunological condition of Litopenaeus vannamei was assessed in semi‐intensive ponds. Earthen ponds were used as experimental units: three with formulated + natural promoted feed + shrimp (T1), three with formulated feed + shrimp (T2), and three with promoted natural feed without shrimp (Control). The dissolved oxygen (DO) levels were optimal for all treatments (≥6 mg L^?1) as well as the pH (8.4–8.6). Total ammonia nitrogen was greater in T2 (0.10 mg L^?1) than T1 (0.07 mg L^?1) and the Control (0.06 mg L^?1). Phytoplankton, zooplankton and benthos were more abundant in T1 and the Control. The promotion of natural feed had a positive effect on all the production parameters of shrimp with an increase of 19.0%, 3.5% and 23.9% in weight gain, survival, and final biomass, respectively; also it was observed a decrease of 13.9% in feed conversion ratio. No differences in haemolymph parameters were observed for nutritional indicators (glucose, cholesterol, proteins, and triglycerides) nor for immunological response (phenoloxidase and prophenoloxidase). The results indicate that the promotion of biotic communities enhances the production parameters of farmed shrimp, without affecting the nutritional and immunological status. Also the water quality was improved by the presence of biota.     

Evaluation of two probiotics used during farm production of white shrimp Litopenaeus vannamei (Crustacea: Decapoda)

This study examined the effects of Bacillus licheniformis strain CIGBC‐232, isolated from the gut of shrimp Litopenaeus vannamei and having antagonistic activity against Vibrio harveyi, on the immunity and larval quality of L. vannamei at various ontogenetic stages, in two separate experiments: (1) PL2 to PL17 exposed to the strain CIGBC‐232 under laboratory conditions (2) zoea I to PL8 exposed to the strain CIGBC‐232 and EPICIN 3W probiotics under farming conditions. The first experiment showed that phenoloxidase, peroxidase and superoxide dismutase activities were improved in animals grown with CIGBC‐232 compared to the untreated control. In tests, the resistance to osmotic stress was also enhanced. During the second experiment, animals treated with CIGBC‐232 exhibited significant (P < 0.05) increases in phenoloxidase activity (30–40% higher in zoea I–II, mysis II–III and PL 2–7) and in the respiratory burst (30% higher from PL 2 to 7) as compared to those animals that received EPICIN 3W. There was no significant difference in the lectins and agglutinins (except in PL1, 6 and 7), in peroxidase and superoxide dismutase activities, in the resistance to osmotic stress, nor in the survival rate among treatments. CIGBC‐232 treatment was able to reduce, the level of presumptive Vibrio spp. with respect to EPICIN 3W treatment in the tank water as well as in animals. At the end of both experiments, the growth of shrimp, i.e. weight and length was increased by CIGBC‐232 treatment. This study showed the probiotic effect of CIGBC‐232, which appeared to have a better probiotic performance than EPICIN 3W treatment.     

TaqMan real-time PCR assay for relative quantification of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia

White spot disease is caused by a highly virulent pathogen, the white spot syndrome virus (WSSV). The disease is usually triggered by changes in environmental parameters causing severe losses to the shrimp industry. This study was undertaken to quantify the relative WSSV load in shrimp exposed to ammonia, using a TaqMan‐based real‐time PCR, and their subsequent susceptibility to WSSV. Shrimp were exposed to different levels of total ammonia nitrogen (TAN) (8.1, 3.8 and 1.1 mg L^?1) for 10 days and challenged with WSSV by feeding WSSV‐positive shrimp. WSSV was detected simultaneously in haemolymph, gills and pereopods at four hours post‐infection. The TaqMan real‐time PCR assay showed a highly dynamic detection limit that spanned over 6 log₁₀ concentrations of DNA and high reproducibility (standard deviation 0.33–1.42) and small correlation of variability (CV) (1.89–3.85%). Shrimp exposed to ammonia had significantly higher (P < 0.01) WSSV load compared to the positive control, which was not exposed to ammonia. Shrimp exposed to 8.1 mg L^?1 of TAN had the highest (P < 0.01) WSSV load in all three organs in comparison with those exposed to 3.8 and 1.1 mg L^?1 of TAN. However, haemolymph had significantly higher (P < 0.01) viral load compared to the gills and pereopods. Results showed that shrimp exposed to ammonia levels as low as 1.1 mg L^?1 (TAN) had increased susceptibility to WSSV.     

Evaluation of three non‐genetically modified soybean cultivars as ingredients and a yeast‐based additive as a supplement in practical diets for Pacific white shrimp Litopenaeus vannamei

Two growth trials and a physiology assessment were conducted to evaluate three non‐genetically modified (GM) soybean cultivars as ingredients in practical diets for Pacific white shrimp, Litopenaeus vannamei. In addition, a commercially available fermented yeast product was evaluated as dietary supplement. For the growth trials (46 and 35 days, respectively, for trials 1 and 2), the basal diet was primarily composed of soybean meal (SBM), fishmeal (FM), whole wheat, corn protein concentrate, poultry meal (PM, pet food grade) and corn starch. Non‐GM cultivars were processed with novel methodologies to produce Navita? ingredients (N1, N2 and N3) which were incorporated at low (L) or high (H) levels into the experimental diets, in partial replacement of FM and full replacement of conventional SBM. The last two formulations incorporated the fermented yeast for a total of nine experimental diets (Table 1 ). Results from the growth trials indicate that shrimp fed diet 5 (HN2) exhibited significantly lower (p < .05) weight gain as compared to shrimp fed diets 1 and 2 (basal and LN1, respectively) in trial 1, as well as compared to animals fed diets 1, 2, 3 and 8 (basal, LN1, HN1 and basal + yeast) in trial 2. The feed conversion ratio significantly increased for shrimp fed diet 5, in contrast with shrimp fed diets 1, 2, 3 and 8 in trials 1 and 2, as well as compared to shrimp fed diets diet 6 (LN3) in trial 2. For the physiological assessment (stress and immune responses), only the effects of diets 1, 3, 8 and 9 (basal, HN1, basal + yeast and HN1 + yeast, respectively) were investigated. Granular cell counts were significantly higher for shrimp fed the yeast‐containing diets. Haemolymph glucose and haemolymph packed cell volume were significantly reduced for shrimp fed diets 3, 8 and 9. No significant differences were observed in total haemocyte counts, hyaline cells counts, semi‐granular cells counts, haemolymph protein, haemocyte phagocytic capacity and haemocyte respiratory burst activity. Results of this work indicate that selective soy breeding technology coupled with novel processing options has the potential to increase the nutritional value of conventional SBM for shrimp feeds. Trends on immune responses were more difficult to elucidate possibly due to the limited length of the feeding trial.     

Effect of recombinant crustacean hyperglycemic hormones rCHH‐B1 and rCHH‐B2 on lipid metabolism in the Pacific white shrimp Litopenaeus vannamei

The crustacean hyperglycemic hormones (CHH) are pleiotropic neuropeptides controlling diverse processes in the life cycle of crustaceans. In the Pacific white shrimp Litopenaeus vannamei, CHH‐B1 and CHH‐B2 are generated by alternative splicing of the same gene, and the recombinant versions of these peptides have shown differential effects on glucose metabolism and osmoregulation in this species. In this work, we explored the role of both recombinant variants, expressed in yeast Pichia pastoris, on lipid metabolism. The results showed that rCHH‐B1 and rCHH‐B2 injections into eyestalk‐ablated shrimp significantly increased the phospholipid and triglyceride levels in haemolymph and restored free fatty acid levels, suggesting that these CHH variants mobilize lipid reserves in shrimp. The effects on lipids were concomitant to its hyperglycemic activity, suggesting that both variants, in addition to their hyperglycemic activity, have hyperlipidemic functions.     

Comparative study of the intracellular superoxide anion production in different penaeid species through the NBT‐reduction assay

The capacity of reactive oxygen intermediates production upon haemocyte stimulation is one of the most important immunoparameter utilized to assess the health status in cultivated shrimps. In the present study, we compared oxidative stress potential, by measuring the superoxide anion production in three penaeid shrimps: two wild Atlantic species, the pink shrimp Farfantepenaeus paulensis and the white shrimp Litopenaeus schmitti and one cultivated Pacific species, the white shrimp, Litopenaeus vannamei, through the nitro‐blue‐tetrazolium‐reduction assay. We also proposed an optimized experimental protocol for this assay, that produces rapid and consistent results with low levels of basal superoxide anion (O₂^?) production by unstimulated haemocytes and high levels of this oxygen radical after cell stimulation. Among the different cell elicitors used (zymosan, laminarin, lipopolysaccharide and phorbol myristate acetate), laminarin (β‐1,3‐glucans – 2 mg mL^?1) was the most potent cell activator for the haemocytes of all three penaeids and we recommend this immunostimulant to routinely evaluate shrimp respiratory burst. In general terms, the most elevated levels of O₂^? production, after cell stimulation with microbial components, were detected in L. schmitti. Interestingly, the stimulation profile of the haemocytes of L. vannamei was more similar to F. paulensis, than to L. schmitti, which is more phylogenetically related.     

Effects of dietary nucleotides on growth,survival and metabolic response in whiteleg shrimp,Litopenaeus vannamei against ammonia stress condition

This trial was conducted to evaluate the effects of nucleotides on growth of whiteleg shrimp, Litopenaeus vannamei, and the survival and metabolic responses to ammonia stress test. Experimental diets were as follows: low fish meal diet (LFMD), and four LFMD test diets, each supplemented with 0.1% guanosine monophosphate (GMP), 0.1% inosine monophosphate (IMP), 0.1% mixture of GMP and IMP and 0.1% mixture of GMP, IMP, uridine monophosphate (UMP) and cytidine monophosphate (CMP). The shrimp specimens (initial body weight: 0.99 ± 0.01 g) were randomly allocated into five groups and fed four times daily for 8‐weeks. After the trial, final body weight was recorded and haemolymph was withdrawn for haematological analysis. The shrimp was then challenged with 70 mg/L ammonia (LC50) for 10 days. Survival and haemolymph of the shrimp were taken after exposure to ammonia. The highest growth performance was observed in the shrimp fed diet supplemented with GMP (p < .05), while survival was not influenced by the test diets in the feeding trial. In the ammonia challenge test, the highest survival was observed in the shrimp fed GMP supplemented diet compared to others. The plasma protein, glucose and cholesterol levels increased in all the treatments while triglycerides level decreased post challenge. Cortisol level recovered at day 10th after the challenge. Shrimps fed with nucleotides diets showed higher protein and glucose level compared to control groups post challenge. In general, nucleotides supplemented in the diet enhanced growth, improved stress resistance while modulating the haemolymph metabolites in L. vannamei under ammonia stress.     

Effects of dietary supplementation of a commercial prebiotic Previda® on survival,growth, immune responses and gut microbiota of Pacific white shrimp,Litopenaeus vannamei

A 35‐day feeding trial was conducted to evaluate growth, bacterial populations of the gastrointestinal tract (GIT) and immune responses of Litopenaeus vannamei fed diets containing the commercial prebiotic Previda^®. Diets were formulated to contain Previda^® at 0, 0.2, 0.5, 1.0 or 1.6 g kg^?1 by weight. At the end of the study, differences in weight gain and survival among treatments were not significant (P > 0.05), but denaturing gradient gel electrophoresis analysis revealed that the microbial communities in the GIT changed significantly with the inclusion of dietary Previda^® at different levels. Previda^® was therefore able to selectively modify the microbial communities in the shrimp's GIT. Although individual bacterial species were not identified, enteric populations in shrimp fed the prebiotic at similar levels of inclusion were genetically similar. In addition, shrimp fed Previda^® at 1.6 g kg^?1 responded significantly (P < 0.05) better immunologically with respect to hemocyte phagocytic capacity, haemolymph protein, hyaline cell counts and haemolymph glucose compared with shrimp fed the basal diet. Although shrimp were not exposed to virulent pathogens in this study, the observed upregulation of some of imm‐une responses upon prebiotic supplementation indicates that an improved outcome of such challenges may be anticipated in Previda^®‐fed shrimp under commercial conditions.     

Evaluation of Agave lechuguilla by‐product crude extract as a feed additive for juvenile shrimp Litopenaeus vannamei

Agave lechuguilla is a succulent plant species, mainly distributed in the northeast of Mexico and south of the United States of America. The main use for this plant is the fibre´s extraction (known as Tampico fibres), resulting in 15% of fibres and 85% of a by‐product waste named guishe. The lechuguilla collectors, normally incinerate the guishe, thus causing environmental contamination. Interestingly, recent studies showed that guishe contains molecules with nutritional properties, such as saponins, flavonoids and sugars. Therefore, in this work, we evaluated the effect of the crude extract of guishe as a feed additive in whiteleg shrimp diets. According to that, MS‐HPLC analysis of the extract showed the presence of saponins such as diosgenin, smilagenin, hecogenin, manogenin, tigogenin hexose, yucagenin, chlorogenin, diosgenin diglucoside and the flavonol, quercetin. After chemical analysis, the crude extract was included into an experimental diet in four levels; 0% (L0%), 0.1% (L0.1%), 0.3% (L0.3%) and 0.6% (L0.6%). Dietary incorporation of the extract was evaluated by zootechnical performance, haemolymph biochemistry, histomorphology and digestive enzyme activity of shrimps. After 5‐week feeding, the L0.3% diet showed significantly higher growth and better feed utilization among treatments. A significant increase in tubule epithelium height and tubule coverage area from hepatopancreas in shrimp under L0.3% diet compared with the control diet suggest an improvement of the health and nutritional status of the shrimp. Inclusion of L0.3% and L0.6% of the crude extract resulted in a reduction in amylase activity, without effect in glucose levels in the haemolymph. Thus, we suggest that lechuguilla guishe crude extract contains nutritional molecules that may be used as a feed additive to promote shrimp productivity.     

Effect of polysaccharides extracts of algae Ulva rigida on growth,antioxidant, immune response and resistance of shrimp,Litopenaeus vannamei against Photobacterium damselae

This study was carried out to investigate the effect of water‐soluble polysaccharides extract of algae Ulva rigida (WPU) as dietary supplement on growth performance, antioxidant enzyme activity, lysozyme and phenoloxidase activity, and resistance of shrimp (Litopenaeus vannamei) subjected to bacterial infection with Photobacterium damselae. Three replicate groups of shrimp (1.0 g) were fed four diets containing four levels, 0 or control, 0.5, 1 and 1.5 g/kg of WPU for 8 weeks over the growth trial. Thereafter, 30 shrimps from each dietary treatment were infected with bacteria P. damselae to evaluate disease resistance of infected shrimp. The results of this study showed that WPU was effective as a growth promoter for L. vannamei. The best growth rate was observed in shrimp fed 1.5 g/kg of WPU diet. Regarding antioxidant defences, the diets supplemented with three levels of WPU stimulated glutathione peroxidase and catalase activates in experimental shrimps. MDA content of L. vannamei‐fed diet containing WPU 1.5 and WPU 1.0 was lower than WPU 0 and WPU 0. 5 diets. Also, lysozyme and phenoloxidase activities of shrimp receiving WPU at 1.0 and 1.5 level were significantly higher than those fed WPU 0 and WPU 0.5 diets. In addition, using WPU extract in all diets decreased mortality in L. vannamei in a dose‐dependent manner after challenge with P. damselae. These results suggest that incorporation of water‐soluble polysaccharides from green algae U. rigida at 1.5 g/kg doses improves growth and antioxidant activity and enhances the immune responses in shrimp L. vannamei.     

Salinity a significant environmental factor for Vibrio harveyi virulence in Fenneropenaeus indicus

The effect of ambient salinity on the haemolymph variables of Fenneropenaeus indicus and its susceptibility to Vibrio harveyi infection under salinity stress has been studied. Adult shrimps were acclimated to 5‰ (hypo osmotic), 25‰ (iso osmotic) and 35‰ (hyper osmotic) salinity levels and the animals were injected with a mid logarithmic culture of V. harveyi at sub lethal level and haemolymph parameters were analysed. Haemolymph proteins, intracellular superoxide anion production, phenoloxidase (PO), alkaline phosphatase (ALP) and acid phosphatase (ACP) activity were found to be at elevated level both at 5‰ and 35‰ post challenge. The haematological responses showed a progressive increase (P < 0.05) up to post challenge day 5 (PCD 5) followed by a considerable decline at all salinities with the lowest being at 35‰. The alterations in the variables were higher in shrimps held at 5‰. However, the V. harveyi infection was severe in animals held at 35‰. The reduction in the parameters could be correlated with the decrease in survival rate of shrimps at 35‰ with a concurrent increase in V. harveyi at this salinity. Multiple regression analysis revealed that ACP (P < 0.001), haemocyte protein HCP (P < 0.001) and PO (P < 0.05) could explain 91% variability in the shrimp survival. These parameters may be used as effective shrimp health indicators. It is evident from the study that ambient salinity alters the haemolymph variables, modulates the virulence in V. harveyi and makes the shrimps more vulnerable to infection at higher salinity. The virulence of V. harveyi is increased at 35‰ salinity as being evidenced from the high mortality at this salinity. The study emphasizes the importance of salinity as an important environmental factor both in terms of host susceptibility and virulence of the pathogen.