Transmission of white spot syndrome virus (WSSV) from Dendronereis spp. (Peters) (Nereididae) to penaeid shrimp

Dendronereis spp. (Peters) (Nereididae) is a common polychaete in shrimp ponds built on intertidal land and is natural food for shrimp in traditionally managed ponds in Indonesia. White spot syndrome virus (WSSV), an important viral pathogen of the shrimp, can replicate in this polychaete (Desrina et al. 2013); therefore, it is a potential propagative vector for virus transmission. The major aim of this study was to determine whether WSSV can be transmitted from naturally infected Dendronereis spp. to specific pathogen‐free (SPF) Pacific white shrimp Litopenaeus vannamei (Boone) through feeding. WSSV was detected in naturally infected Dendronereis spp. and Penaeus monodon Fabricius from a traditional shrimp pond, and the positive animals were used in the current experiment. WSSV‐infected Dendronereis spp. and P. monodon in a pond had a point prevalence of 90% and 80%, respectively, as measured by PCR. WSSV was detected in the head, gills, blood and mid‐body of Dendronereis spp. WSSV from naturally infected Dendronereis spp was transmitted to SPF L. vannamei and subsequently from this shrimp to new naïve‐SPF L. vannamei to cause transient infection. Our findings support the contention that Dendronereis spp, upon feeding, can be a source of WSSV infection of shrimp in ponds.     

Polyculture of Freshwater Angelfish Pterophyllum scalare and Pacific white shrimp Litopenaeus vannamei in low‐salinity water

In this study, we evaluated the production potential of the polyculture of Freshwater Angelfish (Pterophyllum scalare, Cichlidae) and Pacific white shrimp (Litopenaeus vannamei). Two experimental trials were set up. The first trial entailed the use of a randomized design to investigate three treatments options: angelfish monoculture, shrimp monoculture, and fish and shrimp polyculture in 12 experimental 15 m² ponds. In the second trial, we investigated two treatments (polyculture of caged fish and uncaged fish) in 10 experimental units. In trial 1, polyculture negatively affected fish growth and survival. However, fish did not affect shrimp growth and the greatest profit was achieved in polyculture. In trial 2, caging fish improve the growth of both fish and shrimp. An economic analysis showed the greatest profit and benefit cost ratio for caged fish compared with uncaged fish. We conclude that growing P. scalare and L. vannamei together is a better strategy than shrimp monoculture in low‐salinity water. The use of caged fish in such a polyculture operation would enhance productivity and profitability.     

A dual infection by infectious cuticular epithelial necrosis virus and a Chlamydia-like organism in cultured Litopenaeus vannamei (Boone) in Ecuador

During 1996, microscopic examinations of post larvae and juveniles of moribund Litopenaeus vannamei showed multifocal necrosis in the cuticular epithelial tissues. In addition to these severe degenerative alterations in the epithelial cells typical of infectious cuticular epithelial necrosis virus (ICENV), columnar cells of the epithelium displayed small round intracytoplasmic inclusions in the necrotic tissue. Examination by electron microscopy of affected tissues demonstrated prokaryotic organisms in the cytoplasm of epithelial cells delineated by a distinct cytoplasmic vesicle; the prokaryotic organisms were morphologically similar to the genus Chlamydia. The necrotic tissue also showed the presence of particles of ICENV; the double infection by two different organisms in cuticular epithelial cells has not been reported previously. Two distinct stages in the intracellular development of a Chlamydia‐like organism were recognized: (1) pleomorphic elementary bodies (EBs) that were spherical to oval were often observed in the process of division or in forming a common chain of three cells, the cells were surrounded by a rigid cell envelope and the presence of a cap or plaque hexagonally arrayed; (2) the reticular bodies (RBs) were forms often in the process of division. These cells had an electron‐dense cytoplasm and contained a loose network of nuclear fibrils and a more fragile cell envelope. Regardless of the development stages of the Chlamydia‐like organism within the cytoplasmic vesicles, ICENV particles were observed, either dispersed or in clusters, surrounded or inside the vesicles. The potential adverse impact of this dual infection on shrimp culture should be considered, especially in high‐density operations.     

Influence of variation in water temperature on survival,growth and yield of Pacific white shrimp Litopenaeus vannamei in inland ponds for low‐salinity culture

There is considerable interest in the culture of whiteleg shrimp (Litopenaeus vannamei) in inland low‐salinity water in Alabama and other states in the Sunbelt region of the US. However, the growing season is truncated as compared with tropical or subtropical areas where this species is typically cultured, and temperature is thought to be a major factor influencing shrimp production in the US. This study, conducted at Greene Prairie Aquafarm located in west‐central Alabama, considered water temperature patterns on a shrimp farm in different ponds and different years; and sought possible effects of bottom water temperature in ponds on variation in shrimp survival, growth and production. Water temperature at 1.2 m depth in 22 ponds and air temperature were monitored at 1‐hr intervals during the 2012, 2013, 2014 and 2015 growing seasons. Records of stocking rates, survival rates and production were provided by the farm owner. Correlation analysis and linear mixed model analysis of variance were used. Results showed that hourly water temperatures differed among ponds. The range of water temperature in each pond explained 41% of the variance in average final weight of shrimp harvested from each pond. In conclusion, the results suggest that variation in water temperature patterns has considerable influence on shrimp growth and survival in ponds.     

The effects of variation in feed protein level on the culture of white shrimp, Litopenaeus vannamei (Boone) in low-water exchange experimental ponds

The present study was conducted to evaluate the effect of varying dietary protein level on pond water quality and production parameters of white shrimp Litopenaeus vannamei (Boone). Experimental units consisted of nine 400‐m² earthen ponds with a low water exchange. Two treatments were tested: treatment HP consisted of shrimp fed a high‐protein diet (40%) during the whole grow‐out, and treatment LP consisted of the use of a low‐protein diet for the complete farming period. No differences on any of the water quality parameters were observed among treatments. Excellent survival (over 85%) and feed conversion ratios (around 1.6), and acceptable growth (over 12 g) and biomass (from 1721 to 1793 kg ha^?1) were recorded in all experimental ponds. No significant differences in any of the production parameters were found among treatment groups.     

White spot syndrome virus epizootic in cultured Pacific white shrimp Litopenaeus vannamei (Boone) in Taiwan

White spot syndrome virus (WSSV) has caused significant losses in shrimp farms worldwide. Between 2004 and 2006, Pacific white shrimp Litopenaeus vannamei (Boone) were collected from 220 farms in Taiwan to determine the prevalence and impact of WSSV infection on the shrimp farm industry. Polymerase chain reaction (PCR) analysis detected WSSV in shrimp from 26% of farms. Juvenile shrimp farms had the highest infection levels (38%; 19/50 farms) and brooder shrimp farms had the lowest (5%; one of 20 farms). The average extent of infection at each farm was as follows for WSSV‐positive farms: post‐larvae farms, 71%; juvenile farms, 61%; subadult farms, 62%; adult farms, 49%; and brooder farms, 40%. Characteristic white spots, hypertrophied nuclei and basophilic viral inclusion bodies were found in the epithelia of gills and tail fans, appendages, cephalothorax and hepatopancreas, and virions of WSSV were observed. Of shrimp that had WSSV lesions, 100% had lesions on the cephalothorax, 96% in gills and tail fans, 91% on appendages and 17% in the hepatopancreas. WSSV was also detected in copepoda and crustaceans from the shrimp farms. Sequence comparison using the pms146 gene fragment of WSSV showed that isolates from the farms had 99.7–100% nucleotide sequence identity with four strains in the GenBank database – China ( AF332093 ), Taiwan ( AF440570 and U50923 ) and Thailand ( AF369029 ). This is the first broad study of WSSV infection in L. vannamei in Taiwan.     

Experimental white spot syndrome virus challenge of juvenile Litopenaeus vannamei (Boone) at different salinities

In recent years, the shrimp industry has turned to inland freshwater culture as one method to avoid problems such as the introduction of possible vectors of viral pathogens into seawater ponds. Our experiments evaluated susceptibility to white spot syndrome virus (WSSV) in Litopenaeus vannamei held under different salinity regimens. Juvenile L. vannamei that were conditioned at salinities of 35, 25, 15, 5 and 2 g L⁻¹ were challenged with WSSV. In order to assess the severity of white spot disease, histological analysis and nested polymerase chain reaction (PCR) tests were carried out on the challenged shrimp every 4 h after 48 h post challenge. The results indicated that significantly more severe infections resulted at 15‰ than at other salinities. Mortality could not be compared due to the sampling design and because severe WSSV infections occurred in all test groups such that few shrimp remained alive in each challenged group at the end of the test. Despite this, the results suggest that salinity may affect the course and outcome of WSSV infections.     

Litopenaeus vannamei (Boone) post-larval survival related to age, temperature, pH and ammonium concentration

Transport of post‐larvae shrimp used in aquaculture is an important element of successful cultivation because of the potential for stress during stocking procedures. To find optimum transport conditions, several bioassays were performed in the laboratory to evaluate survival of whiteleg shrimp Litopenaeus vannamei 5–30‐day‐old postlarvae under conditions similar to those encountered during transport from the hatchery to nursery and shrimp ponds. Postlarvae were exposed for 4 h to different temperatures and pH levels ammonia concentrations. Survival was significantly reduced after a 4 h exposure to pH 9 and was inversely related to temperature with or without 7 mg L^?1 of ammonia. The 15‐ and 20‐day‐old postlarvae had higher survival rates than other ages. The lowest survival occurred in alkali conditions (pH 9), with 7 mg L^?1ammonia at 30 and 32°C. To assure optimal survival of postlarvae during transfer from the hatchery to the nursery and shrimp ponds, we recommend temperatures below 28°C, pH no higher than 8, no ammonia and post‐larval age at least 15 days.     

Infection of IHHN virus in two species of cultured penaeoid shrimp Litopenaeus vannamei (Boone) and Litopenaeus stylirostris (Stimpson) in Ecuador during El Niño 1997–98

Infectious hypodermal and haematopoietic necrosis virus (IHHNV) was first noted in blue shrimp Litopenaeus stylirostris (Stimpson) in mid-1981. Since that time, at least 12 species of penaeoid shrimp have been reported to be infected with IHHNV. Pacific white shrimp Litopenaeus vannamei (Boone) represents a shrimp species highly refractory to the disease, whereas L. stylirostris was highly susceptible to the disease. Since the beginning of the shrimp farming industry in Ecuador, viral diseases have been observed in L. vannamei and L. stylirostris. Of these, L. vannamei represents ≈ 80% of cultured shrimp. Histopathology, ultrastructure and in situ DNA hybridization confirm the presence and assess the prevalence of IHHNV in pond-reared shrimp, and especially in abnormally small animals of both species. Although IHHNV may be considered enzootic in cultured L. vannamei in Ecuador, we did not find high prevalence (Cowdry A bodies) in specimens of diseased pond shrimp before 1996. From that time to 1998, a higher prevalence of IHHNV has been observed in both species. The epizootic of the IHHNV disease has been related to the oceanographic and climatological conditions caused by El Niño. In addition, it has been suggested that large quantities of wild shrimp post-larvae of both species that were stocked in shrimp farms, infected as latent carriers in 1997, from which the virus could spread to a larger population of these shrimp in 1998.     

Pond production of Pacific white shrimp,Litopenaeus vannamei,fed grain distillers dried yeast

The objective of this study was to demonstrate the feasibility of four diets formulated to contain increasing levels (0, 50, 100 and 150 g kg^?1 of diet) of grain distillers dried yeast (GDDY) in production diets for Litopenaeus vannamei, reared in outdoor tanks or production ponds. The production pond trial was carried out in 16, 0.1‐ha ponds using four replicates per diet. Juvenile shrimp (38.1 ± 4.26 mg, initial weight) were stocked at 30 shrimp m^?2 for a 16‐week period. The same four diets and a commercial reference diet were offered to shrimp maintained in outdoor tanks over a 12‐week period. A total of 20 tanks were stocked with juvenile shrimp (3.05 ± 0.22 g, initial weight) obtained from production ponds at a density of 30 shrimp per tank (40 shrimp m^?2). At the conclusion of these trials, mean final weight ranged from 19.77 to 23.05 g, yield ranged between 4760 and 5606 kg ha^?1, survival ranged from 69.6% to 89.4%, and feed conversion ratio (FCR) was between 1.02 and 1.23. Shrimp reared in the outdoor tanks confirmed the results of the pond trial. Mean final weight ranged between 18.12 and 18.97 g, survival ranged from 93.3% to 98.3%, and FCR was between 1.25 and 1.29. In both trials, there were no significant differences regarding mean final weight, FCR and survival among dietary treatments. Based on this study, GDDY up to 150 g kg^?1 of diet can be used in L. vannamei commercial feed formulation.     

A novel micro‐organism for removing excess ammonia‐N in seawater ponds and the effect of Cobetia amphilecti on the growth and immune parameters of Litopenaeus vannamei

To investigate the feasibility of using micro‐organisms for ammonia‐N removal, six strains were isolated from Chinese white shrimp, Fenneropenaeus chinensis, seawater culture ponds in Dongying (Shandong, China). Of these, strain DY‐01, which exhibited the highest capacity to degrade ammonia‐N, removed 61.7% of the total ammonia‐N (50 mg/L) in 8 hr. An investigation of the factors affecting the removal efficiency indicated the optimum conditions to be 30°C, pH 8.0, and a salinity level of 30 g/L; 16S rDNA gene sequencing and biochemical analysis identified strain DY‐01 as Cobetia amphilecti, which has not previously been reported to degrade ammonia‐N. This strain also boosted the growth of Pacific white shrimp, Litopenaeus vannamei (p < .05), at a concentration of 10⁷ CFU/mL, with no harmful effects on the shrimp's immune system. This study has thus identified a novel aerobic nitrifying bacterium that is potentially an excellent candidate for improving the water quality in mariculture ponds.     

Effects of Inorganic and Organic Fertilization on Physicochemical Parameters,Bacterial Concentrations,and Shrimp Growth in Litopenaeus vannamei Cultures with Zero Water Exchange

To identify ways to improve water quality and shrimp production in closed systems, two parallel experiments (one in tanks and one in ponds) were conducted using Pacific white shrimp, Litopenaeus vannamei, cultures. In both experiments, the effects of inorganic (Nutrilake®) and organic (molasses) fertilization on physicochemical parameters, bacterial concentrations, and shrimp performance under zero water exchange were evaluated. Fertilization with both molasses and Nutrilake enhanced the feed conversion rate, as well as shrimp survival and production. In tanks, the shrimp survival and production rates were highest in the molasses treatment, but this effect was not observed in ponds. In ponds, fertilization with Nutrilake increased nitrogen and phosphorus concentrations more than did the controls and molasses treatments toward the end of the experiment. In tanks, fertilization with molasses reduced ammonia concentrations toward the end of the experiment, but the same effect was not observed in ponds. In ponds, fertilization reduced the proportion of Vibrio spp. bacteria, which most likely reduced the incidence of disease from these potentially pathogenic organisms. In both culture systems, fertilization increased the proportion of Bacillus spp., which most likely enhanced food availability.     

Biochemical changes and tissue distribution of Enterocytozoon hepatopenaei (EHP) in naturally and experimentally EHP‐infected whiteleg shrimp,Litopenaeus vannamei (Boone, 1931), in India

Stunted growth in pond‐reared Litopenaeus vannamei was observed in different farms located in Tamil Nadu and Andhra Pradesh, India. No mortality was associated with stunted growth. PCR assay on these samples revealed the presence of Enterocytozoon hepatopenaei (EHP) in stunted shrimp. Tissue distribution of EHP in naturally and experimentally infected shrimp was studied by PCR and histology. Histological examination revealed the presence of EHP in hepatopancreas and gut, but not in other organs. The PCR assay revealed the presence of EHP in all the organs tested in both naturally and experimentally infected shrimp. Healthy shrimp were challenged with E. hepatopenaei by intramuscular injection and oral route, and no mortality was observed in both routes after 30 days post‐challenge. Different developmental stages of the microsporidian parasite were observed in the hepatopancreatic epithelial cells. Biochemical parameters such as total protein, albumin, aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase were measured in the haemolymph of naturally and experimentally EHP‐infected shrimp. All biochemical parameters mentioned were found to be significantly higher in EHP‐infected shrimp when compared to normal shrimp. This is the first report relating AST and ALT levels to EHP infection in naturally and experimentally infected shrimp.     

Impact of yellow head virus outbreaks in the whiteleg shrimp,Penaeus vannamei (Boone), in Thailand

Yellow head virus (YHV) is known as a major pathogen in the black tiger shrimp, Penaeus (Penaeus) monodon. It can also cause serious mortality in farmed whiteleg shrimp, Penaeus (Litopenaeus) vannamei. However, there is no published information on the economic and/or production impact of the disease in P. vannamei. Shrimp with gross signs of YHV disease (faded body colour and 60–70% mortality) were observed in 20 study farms rearing P. vannamei in the central part of Thailand from the end of 2007 through early 2008. The estimated economic loss for these farms according to the Thai Animal Aquaculture Association was approximately US$3 million. Detailed sequence analysis of RT‐PCR amplicons from shrimp in all the study ponds revealed the presence of YHV Type 1b (YHV‐1b) alone (characterized by a 162‐bp deletion in the ORF3 region encoding the structural gene for gp116) and the absence of YHV Type 1a (YHV‐1a), the original YHV type reported from Thailand. Despite the large 162‐bp deletion (= 54 deduced amino acids) in the gp116 structural gene, histopathology of YHV‐1b infections was identical to that of YHV‐1a infections, and electron microscopy revealed that YHV‐1b virions were morphologically indistinguishable from those previously reported for YHV‐1a. In addition, an existing commercial RT‐PCR detection kit and an immunochromatographic test strip for the detection of YHV were proven to have been valid tests for both YHV‐1b and YHV‐1a. The source of the virus for these outbreaks was unlikely to have been the post‐larvae used to stock the ponds, as they were derived from domesticated specific pathogen‐free stocks free of YHV. Thus, it is possible that they originated from an unknown, natural reservoir.     

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.     

Nematopsis marinus n. sp., a new septate gregarine from cultured penaeoid shrimp Litopenaeus vannamei (Boone), in Ecuador

The septate gregarine Nematopsis marinus n. sp. (Apicomplexa: Cephaline) heavily infected the midgut of cultured Pacific white shrimp Litopenaeus vannamei (Boone) in Ecuador. It is morphologically similar to other species of the genera Nematopsis, but it can be distinguished from them, by having gamonts with a prominent hemispherical protomerite that contained numerous refractile granules and unusual strong gliding movement. There is evidence that shrimp acquired the infection in the ponds, as larval or postlarval stages do not showed infection. Juveniles and adult shrimp had a prevalence and intensity of infection ranging from 50% to 80% and 10 to > 5000 parasites respectively. When voided from the gut, the gregarine keep alive in seawater. This gregarine have been associated with the marine environment and there are no records of this species in low salinity waters or freshwater. Results suggest that N. marinus could have most of the life cycle of the species within the host L. vannamei.     

Sperm capacitation of the shrimp Litopenaeus vannamei

Litopenaeus vannamei is one of the most important species of farmed shrimp. The females have an ‘open’ thelycum. Mating is accomplished by attaching the male spermatophore onto the surface of the thelycum 4–6 h before spawning. During this period, sperm may have to undergo morphological changes associated with a capacitation process that has been described for other shrimp species. The objective of this research was to extend research on sperm capacitation in L. vannamei by ultrastructural and biochemical means. The sperm of L. vannamei were divided into those freshly prepared from the spermatophore (S‐sperm), extracted from the male gonopores, and those extracted from the female thelycum (T‐sperm). Under transmission electron microscopy, ultrastructural differences were detected between the S‐ and the T‐sperm in the nuclear material, the filamentous meshwork and the cytoplasmic particles. Under scanning electron microscopy, the difference was observed in the cap and spike regions. Immunofluorescence using confocal microscopy to detect tyrosine phosphorylated proteins revealed different distribution patterns between S‐ and T‐sperm. The location of phosphorylation activity changed from the spike in S‐sperm, to the filamentous meshwork in the T‐sperm. These morphological and biochemical changes confirm that capacitation of L. vannamei sperm takes place following mating.     

Effects of water hyacinth (Eichhornia crassipes) leaf protein concentrate as soybean protein replacement in white shrimp Litopenaeus vannamei (Boone) postlarvae diet

The potential of water hyacinth leaf protein concentrate (WHLPC) as an alternative protein source to soybean meal in formulated diets for the white shrimp Litopenaeus vannamei (Boone) postlarvae, was assessed. Isonitrogenous and isolipidic diets were formulated containing 40% protein and 8% lipid. WHLPC replaced 0%, 25%, 50% and 75% of soybean meal protein in the formulated diets. Twenty postlarval Litopenaeus vannamei weighing 5.0 ± 0.00032 mg were randomly assigned to twelve 30‐L aquarium tanks and were fed experimental diets for 70 days. Results show that the overall biological performance of shrimp in terms of specific growth rate (SGR), protein efficiency ratio (PER), feed efficiency (FE) and survival in treatment receiving the 25% WHLPC replacement is comparable to that of the control diet. Replacement levels beyond 25% exhibited significant depression in terms of growth performance of the shrimp. Collectively, these findings indicate that WHLPC can replace 25% of soybean meal protein in the formulated diets of L. vannamei.