Edgar C. Amar

Acute hepatopancreatic necrosis disease (AHPND) outbreaks in Penaeus vannamei and P. monodon cultured in the Philippines

Acute hepatopancreatic necrosis disease (AHPND) has recently emerged as a serious disease of cultured shrimp. It has also been described as early mortality syndrome (EMS) due to mass mortalities occurring within 20 to 30 d after stocking of ponds with postlarvae. Here, Penaeus vannamei and Penaeus monodon from shrimp farms in the Philippines were examined for the toxin-producing strain of Vibrio parahaemolyticus due to AHPND-like symptoms occurring in marketable size shrimp. In the P. vannamei, histology revealed typical AHPND pathology, such as sloughing of undifferentiated cells in the hepatopancreatic tubule epithelium. Analysis using the IQ2000 AHPND/EMS Toxin 1 PCR test generated 218 bp and 432 bp amplicons confirmative of the toxin-producing strain of V. parahaemolyticus among shrimp sampled from 8 of 9 ponds. In the P. monodon, histology revealed massive sloughing of undifferentiated cells of the hepatopancreatic tubule epithelium in the absence of basophilic bacterial cells. PCR testing generated the 2 amplicons confirmatory for AHPND among shrimp sampled from 5 of 7 ponds. This study confirms the presence of AHPND in P. vannamei and P. monodon farmed in the Philippines and suggests that the disease can also impact late-stage juvenile shrimp.

Poly-β-hydroxybutyrate (PHB) accumulating Bacillus spp. improve the survival, growth and robustness of Penaeus monodon (Fabricius, 1798) postlarvae.

Low larval survival resulting from suboptimal culture conditions and luminous vibriosis poses a major problem for the larviculture of penaeid shrimp. In this study, a poly-β-hydroxybutyrate (PHB) accumulating mixed bacterial culture (mBC; 48.5% PHB on cell dry weight) and two PHB accumulating bacterial isolates, Bacillus sp. JL47 (54.7% PHB on cell dry weight) and Bacillus sp. JL1 (45.5% PHB on cell dry weight), were obtained from a Philippine shrimp culture pond and investigated for their capacity to improve growth, survival and robustness of Penaeus monodon postlarvae (PL). Shrimp PL1 and shrimp PL30 were provided with the PHB containing bacterial cultures in the feed for 30 days followed by, respectively, a challenge with pathogenic Vibrio campbellii and exposure to a lethal dose of ammonia. Prior to the pathogenic challenge or ammonia stress, growth and survival were higher for shrimp receiving the PHB accumulating bacteria as compared to shrimp receiving diets without bacterial additions. After exposure to the pathogenic challenge the shrimp fed PHB accumulating bacteria showed a higher survival as compared to non-treated shrimp, suggesting an increase in robustness for the shrimp. Similar effects were observed when shrimp PL30 were provided with the PHB accumulating bacterial cultures during a challenge with pathogenic V. campbellii through the water. The survival of shrimp exposed to lethal ammonia stress showed no significant difference between PHB accumulating bacteria-fed shrimp and non-PHB treated shrimp. The data illustrate that bacilli capable of accumulating PHB can provide beneficial effects to P. monodon post-larvae during culture in terms of growth performance, survival and resistance against pathogenic infection and ammonia stress. Further investigations are required to verify the PHB effect of the bacterial cultures on the shrimp.

A probiotic Bacillus strain containing amorphous poly-beta-hydroxybutyrate (PHB) stimulates the innate immune response of Penaeus monodon postlarvae

Abstract In this study, the PHB‐accumulating Bacillus sp. JL47 strain (capable of accumulating 55% PHB on cell dry weight) was investigated for its effects on the immune response of giant tiger shrimp (Penaeus monodon) postlarvae (PL) before and after the Vibrio campbellii challenge. Briefly, shrimp PL were cultured and fed with Artemia nauplii enriched with Bacillus sp. JL47. Shrimp receiving the Artemia nauplii without JL47 enrichment were used as control. After 15 days of feeding, the shrimp were challenged with pathogenic V. campbellii LMG 21363 at 106 cells mL−1 by immersion. Relative expression of the immune related genes encoding for prophenoloxidase (proPO), transglutaminase (TGase) and heat shock protein 70 (Hsp70) in the shrimp were measured before (0 h) and after (3, 6, 9, 12, 24 h) the Vibrio challenge by quantitative real‐time PCR using &bgr;‐actin as the reference gene. The expressions of TGase and proPO were significantly up‐regulated (p < 0.05) within 9 h and 12 h, respectively after challenge in shrimp receiving the Bacillus sp. JL47 as compared to the challenged and non‐challenged controls. Hsp70 expression was significantly increased (p < 0.05) at 3 h post‐challenge in all challenged shrimp. Interestingly, proPO and TGase genes were significantly up‐regulated (p < 0.05) in Bacillus sp. JL47 treated shrimp even before the Vibrio challenge was applied. No up‐regulation in the Hsp70 gene, however, was observed under these conditions. The data suggest that the protective effect of the PHB‐accumulating Bacillus sp. JL47 in shrimp was due to its capacity to stimulate the innate immune related genes of the shrimp, specifically the proPO and TGase genes. The application of probiotic Bacillus species, capable of accumulating a significant amount of PHB, is suggested as potential immunostimulatory strategy for aquaculture. HighlightsThe PHB‐accumulating Bacillus sp. JL47 strain stimulates the proPO and TGase genes of P. monodon PL.Stimulation of proPO and TGase in shrimp was observed before and after the V. campbellii challenge.Stimulation of Hsp70 gene was observed in all challenged shrimp 3 h after the V. campbellii exposure.Use of probiotic Bacillus containing amorphous PHB is a promising immunostimulatory strategy.