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.     

Fortification of Bread Wheat Using Synthesized Zn-Glycine and Zn-Alanine Chelates in Comparison with ZnSO4 in a Calcareous Soil

ABSTRACT

Calcareous soils typically suffer from zinc deficiency and zinc sulfate is incorporated in many cultivated soils. Utilization of ZnSO₄ has some kinds of interaction with soil particles and organic matter. In this study, the efficacy of two znic(Zn)-amino acid chelates (Zn-ACs) i.e., Zn-alanine (Zn-Ala) and Zn-glycine (Zn-Gly) on wheat (Triticum aestivum, cv. N91-8) growth characteristics and zinc concentration in wheat was examined under greenhouse conditions and compared to the a commercial ZnSO₄. Results showed that Zn-Ala and Zn-Gly significantly increased the dry weight and shoot length of wheat in comparison to ZnSO₄ treatment. Soil application of Zn-Amino acid chelates proved to be the most influential source of zinc in increasing wheat growth and yield indices. Number of fertile spikelet and grain yield increased significantly respectively compared to ZnSO₄ treatment. Zn concentration and protein content of wheat grain in Zn-ACs treatment was significantly higher than the ZnSO₄ treatment. Soil application of Zn-ACs caused a significant decrease in the grain phytic acid (PA) concentration and also phytic acid to zinc molar ratio in comparison with ZnSO₄ treatment. According to the results, Zn-ACs could be utilized as a zinc fertilizer source for improving the zinc bioavailability in wheat.     

Evaluation of limb autotomy as a promising strategy to improve production performances of mud crab (Scylla olivacea) in the soft‐shell farming system

Soft‐shell crab, a newly moulted crab having non‐calcified and hydrated soft exoskeleton, farming has been increasing tremendously in recent years throughout the world due to its high market price and consumer preferences. However, high aggressiveness and cannibalistic behaviour of mud crab limit its culture to an individual chamber system which ultimately makes the soft‐shell mud crab farming space and labour‐intensive. Therefore, the aim of this study was to improve the efficiency of soft‐shell crab production by autotomy of the claw and/or walking legs to reduce the moulting duration and intensify the culture system through double stocking in the existing individual chamber culture system. To achieve our objectives, a series of experiment were conducted at both indoor and on‐farm levels by removing the claw and/or walking legs of Scylla olivacea for two consecutive moulting cycles. Results showed that both claw ablation significantly (p < .01) shortened the moulting duration and increased the carapace width, weight gain, specific growth rate, feeding efficiency and yield parameter compared to the non‐ablated crabs more prominently at 1st moulting than the 2nd moulting in both indoor and on‐farm trials. Ablation of all walking legs while keeping the claw intact did not significantly influence (p > .05) the moulting duration and carapace width, but significantly (p < .01) improved the specific growth rate, weight gain, feeding efficiency and yield parameters at 1st moulting compared to the non‐ablation and unilateral three walking leg ablation groups. However, ablation of all appendages (both claw and walking legs) showed significant (p < .01) improvements of carapace width, weight gain, specific growth rate, feeding efficiency and yield parameters, and shortened the moulting duration up to 23 days from 34 days during 1st moulting and up to 27 days from 40 days at 2nd moulting. The survivals of different autotomized mud crabs were statistically non‐significant (p > .05) to the non‐autotomized crabs in various indoor and on‐farm trials. Besides improving the production performances, ablation of bilateral claw and all appendages allowed to double the stocking density in the existing individual chamber‐based soft‐shell crab farming that enhanced the production about threefold higher than the existing individual chamber‐based culture system. Finally, our results suggest that autotomy could be a promising technology for the commercial farming of soft‐shell mud crab in near future.     

SnToxA,SnTox1, and SnTox3 originated in Parastagonospora nodorum in the Fertile Crescent

The centre of origin of the globally distributed wheat pathogen Parastagonospora nodorum has remained uncertain because only a small number of isolates from the Fertile Crescent were included in earlier population genetic and phylogeographic studies. We isolated and genetically analysed 193 P. nodorum strains from three naturally infected wheat fields distributed across Iran using 11 neutral microsatellite loci. Compared to previous studies that included populations from North America, Europe, Africa, Australia, and China, the populations from Iran had the highest genetic diversity globally and also exhibited greater population structure over smaller spatial scales, patterns typically associated with the centre of origin of a species. Genes encoding the necrotrophic effectors SnToxA, SnTox1, and SnTox3 were found at a high frequency in the Iranian population. By sequencing 96 randomly chosen Iranian strains, we detected new alleles for all three effector genes. Analysis of allele diversity showed that all three effector genes had higher diversity in Iran than in any population included in previous studies, with Iran acting as a hub for the effector diversity that was found in other global populations. Taken together, these findings support the hypothesis that P. nodorum originated either within or nearby the Fertile Crescent with a genome that already encoded all three necrotrophic effectors during its emergence as a pathogen on wheat. Our findings also suggest that P. nodorum was the original source of the ToxA genes discovered in the wheat pathogens Phaeosphaeria avenaria f. sp. tritici 1, Pyrenophora tritici-repentis, and Bipolaris sorokiniana.     

An Electrothermochromic Fabric Prepared by Electrodeposition of Polypyrrole on Single Side

An electrothermochromic fabric triggered by electric voltages was prepared by combining the electric heating feature of conductive polypyrrole (PPy) and the temperature-response color-changing feature of thermochromic inks. PPy was deposited selectively on the carbon-coated side of the sailcloth fabric via electrochemical polymerization. Thermochromic inks were painted on the reverse side with white color. The obtained fabric could be heated to about 48 °C and change its color significantly under an applied voltage of 3 V.     

Recent update on the prevalence of Vibrio species among cultured grouper in Peninsular Malaysia

Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene.     

Physiological response,blood chemistry profile and mucus secretion of red sea bream (<Emphasis Type="Italic">Pagrus major</Emphasis>) fed diets supplemented with <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> under low salinity stress

Environmental stressors caused by inadequate aquaculture management strategies suppress the immune response of fish and make them more susceptible to diseases. Therefore, efforts have been made to relieve stress in fish by using various functional feed additives in the diet, including probiotics. The present work evaluates the effects of Lactobacillus rhamnosus (LR) on physiological stress response, blood chemistry and mucus secretion of red sea bream (Pagrus major) under low salinity stress. Fish were fed four diets supplemented with LR at [0 (LR0), 1 × 10² (LR1), 1 × 10⁴ (LR2) and 1 × 10⁶ (LR3) cells g^?1] for 56 days. Before stress, blood cortisol, urea nitrogen (BUN) and total bilirubin (T-BIL) showed no significant difference (P > 0.05), whereas plasma glucose and triglyceride (TG) of fish-fed LR2 and LR3 diets were significantly lower (P < 0.05) than those of the other groups. Plasma total cholesterol (T-CHO) of fish-fed LR3 diet was significantly (P < 0.05) lower than that of the other groups. Furthermore, total plasma protein, mucus myeloperoxidase activity and the amount of mucus secretion were significantly enhanced in LR-supplemented groups when compared with the control group (P < 0.05). After the application of the low salinity stress test, plasma cortisol, glucose, T-CHO and TG contents in all groups showed an increased trend significantly (P < 0.01) compared to the fish before the stress challenge. However, plasma total protein and the amount of secreted mucus showed a decreased trend in all groups. On the other hand, BUN, T-BIL and mucus myeloperoxidase activity showed no significant difference after exposure to the low salinity stress (P > 0.05). In addition, the fish that received LR-supplemented diets showed significantly higher tolerance against low salinity stress than the fish-fed LR-free diet (P < 0.05). The physiological status and the detected immune responses, including total plasma protein and mucus myeloperoxidase activity in red sea bream, will provide a more comprehensive outlook of the effects of probiotics to relieve stress in fish.     

Nitrogen Fertilizer Management Practices to Reduce N<Subscript>2</Subscript>O Emissions from Irrigated Processing Potato in Manitoba

Nitrogen fertilizer practices affect nitrous oxide (N₂O) emissions from agricultural soils. The “4R” nutrient stewardship framework of using N fertilizer at the right rate, right source, right placement and right time can reduce N₂O emissions while maintaining or improving yield of field crops, but understanding of how the various factors affect N₂O emissions from irrigated processing potato is lacking. We examined the effects of selected 4R practices on emissions, using results from two irrigated processing potato studies each conducted in 2011 and 2012 in Manitoba, Canada. Experiment 1 examined combinations of source (urea, ESN), placement (pre-plant incorporation [PPI], banding), and rate (100 and 200 kg N ha^-1) on a clay loam soil. Experiment 2 examined timing and source treatment combinations (urea PPI, ESN PPI, urea split, urea split/fertigation) on a loamy fine sandy soil. For Experiment 1, use of ESN at 200 kg ha^-1 did not reduce area-, yield- and applied fertilizer N- based N₂O emissions compared to urea at 200 kg ha^-1, irrespective of placement. Emissions from pre-plant banding ESN at 200 kg ha^?1, however, were 32% lower than from PPI ESN. For Experiment 2, compared to single pre-plant urea application, fertigation simulated by in-season application of urea ammonium nitrate (UAN) gave lower area-, yield- and applied fertilizer N- based emissions. Split urea ( \( \raisebox{1ex}{$2$}\!\left/ \!\raisebox{-1ex}{$3$}\right. \) pre-plant, \( \raisebox{1ex}{$1$}\!\left/ \!\raisebox{-1ex}{$3$}\right. \) hilling) also reduced area- and yield- based N₂O emissions compared to single pre-plant urea application. Emissions were generally lower at the site with loamy fine sandy soil than the site with clay loam soil. These results demonstrate that combinations of “4R” practices rather than source alone are best to achieve reductions in N₂O emissions from irrigated potato production.