Characterization of Aquimarina hainanensis isolated from diseased mud crab Scylla serrata larvae in a hatchery

Mass mortality due to necrosis signs occurred in hatchery-reared zoea stage larvae of the mud crab Scylla serrata in Okinawa, Japan, and a causative bacterium was isolated. In this study, we identified and characterized the bacterium by genome analysis, biochemical properties and pathogenicity. The bacterium was a Gram-negative, non-motile, long rod, forming yellow colonies on a marine agar plate. It grew at 20–33°C (not at 37°C) and degraded chitin and gelatin. Phylogenetic analysis of the 16S rRNA gene sequence identified the bacterium as Aquimarina hainanensis. Genome sequence data obtained from Illumina MiSeq generated 29 contigs with 3.56 Mbp in total length and a G + C content of 32.5%. The predicted 16 chitinase genes, as putative virulence factors, had certain homologies with those of genus Aquimarina. Experimental infection with the bacterium conducted on larvae of four crustacean species, brine shrimp Artemia franciscana, freshwater shrimp Caridina multidentata, swimming crab Portunus trituberculatus and mud crab S. serrata, revealed that this bacterium was highly virulent to these species. The present study suggests that the bacterium caused mass mortality in mud crab seed production was A. hainanensis and can be widely pathogenic to crustaceans.     

Antiparasitic activity of the medicinal plant Dillenia suffruticosa against the marine leech Zeylanicobdella arugamensis (Hirudinea) and its phytochemical composition

Zeylanicobdella arugamensis (Annelida: Hirudinea), a marine parasitic leech, is currently affecting different species of cultured groupers, hybrid groupers, snappers and sea bass in Malaysia. Dillenia suffruticosa (Dilleniaceae), a medicinal plant found in Sabah, has been selected in our experiment to kill the leeches as a natural control method. The leech‐infested hybrid groupers were collected from aquaculture facilities, and the isolated leeches were challenged against methanol extract of D. suffruticosa leaves. The experiment was carried out using various concentrations of the extracts such as 25, 50 and 100 mg/ml. The methanol extract showed significant antiparasitic activity against Z. arugamensis with 100% mortality at a concentration of 100 mg/ml in 14.39 ± 3.75 min., followed by 50 and 25 mg/ml in 32.97 ± 9.29 and 41.77 ± 5.40 min., respectively. The phytochemical composition of the extract was determined using GC‐MS analysis to understand the nature of the principal compounds responsible for its antiparasitic properties. The leaves of D. suffruticosa demonstrated the presence of different bioactive compounds of various natures with varying percentages. Thus, it could be revealed that the methanol extract of D. suffruticosa mainly contains vital phytochemical compounds and showed an effective antiparasitic activity against the harmful leeches of hybrid groupers.     

Physical properties of tsunami-affected soils in Aceh, Indonesia: 2½ years after the tsunami

On 26 December 2004, a tsunami caused extensive loss of life, damaged property and degraded agricultural land in the province of Aceh, Indonesia. While some of the associated soil chemical changes have been documented, information on soil physical properties is sparse. The objective of this study was to quantify physical properties of some tsunami-affected upland agricultural soils in Aceh, Indonesia. Soil was sampled approximately 21/2 years after the tsunami, from the 0–0.1 m, 0.1–0.3 m and 0.3–0.5 m depths in four sites in the villages of Kling Cot Aroun in Aceh Besar sub-district, Kuta Kruen in Aceh Utara sub-district, Udjong Blang Mesjid in Bireuen sub-district and Meue in Pidie Jaya sub-district on the east coast of Aceh. These sites were located within 1 km from the sea at elevations ranging from 0 to 5 m ASL. The soils were Ultisols except for Meue, which was an Entisol. Soil properties measured were bulk density, structural stability and particle size distribution. Soil water retention, pore-size distribution and saturated hydraulic conductivity were estimated by inserting the values of bulk density, clay, sand and silt contents into pedotransfer functions from the literature. The analyses conducted during this study did not permit us to ascertain what proportion of the soil particles were of tsunami-origin. Nonetheless, deposition of finer-textured material may have occurred in two of the sites. In comparison with the greyish-white, coarse textured soil in the rest of the profile, a finer-textured yellow horizon was present in the lower slopes of the Udjong Blang Mesjid site. At Meue, clay and silt contents were higher in the surface 0.3 m than in the 0.3–0.5 m depth, although a distinct horizon was absent. Particle size distribution in all sites was dominated by the sand fraction, although clay and silt contents were relatively high (20–30 g 100 g^− 1) at Kuta Kruen. Among the sand fractions, fine sand (0.02–0.25 mm) was highest at Kling Cot Aroun, Kuta Kruen and in the “yellow horizon” at Udjong Blang Mesjid, making them more prone to hardsetting and compaction after intensive tillage. Soil compaction was present in all sites with that in the “yellow horizon” at Udjong Blang Mesjid being highest. The relatively low porosity in this layer may be beneficial, as it is likely to reduce the high rates of water drainage and nutrient leaching in this sandy soil. The more compacted soils were characterised by higher numbers of micropores (r, pore radius < 4.3 μm), lower water retention at saturation, smaller numbers of macropores (r > 14.3 μm), lower hydraulic conductivity and intensive gleying, indicating frequent waterlogging. The soils in all depths from Kling Cot Aroun and the “yellow horizon” at Udjong Blang Mesjid were very dispersive, that at Meue moderately dispersive in the 0.3–0.5 m depth but stable in the 0–0.1 m depth, and at Kuta Kruen very stable in all depths. Soil physical degradation was a feature of the soils examined, and its amelioration will be the key to improving and sustaining crop yields in these soils. Possible management interventions include organic amendments such as compost or manure, and minimum tillage options such permanent beds or zero tillage with retention of crop residues as in situ mulch together with suitable cover crops.     

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.     

Isolation and Characterization of Large Marine Bacteriophage (Myoviridae), VhKM4 Infecting Vibrio harveyi

Abstract

The causative agent responsible for vibriosis in tropical fish aquaculture, Vibrio harveyi, has become a major bacterial pathogen. Studies suggest that this bacterium has developed resistance to antibiotics commonly used in aquaculture. In view of this situation and the requirement for the proposed postantibiotic era, bacteriophage therapy seems to be a promising control strategy for fish vibriosis. In this study, a lytic Vibrio phage VhKM4 belonging to a member of large, marine Myoviridae was successfully isolated. It exhibited bacteriolysis to both V. harveyi VHJR7 and V. parahaemolyticus ATCC 17802. The latent period of the VhKM4 phage was recorded at 60 min. It also recorded average burst size of approximately 52 plaque-forming units per infected cell. A strong bacteriolytic activity at low multiplicity of infection of 0.01 indicates the effectiveness of this large marine myovirid against fish pathogenic strain of V. harveyi VHJR7.

Received June 16, 2016; accepted October 7, 2016 Published online February 6, 2017