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.     

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.     

Assessment of the Antibacterial Effect of Chitosan Coated with Heracleum persicum Oil on Rainbow Trout Quality

Today, due to the rapid spoilage of fish, the use of natural preservatives is a priority over those of synthetic varieties. Also, the natural antimicrobial effects of essential oils can help to increase shelf life. There are very few studies concerning the use of essential oils in this regard. In the present study, the antibacterial effect of chitosan coated with Heracleum persicum oil was investigated on the quality of rainbow trout. The control and the coated fish samples were analyzed periodically by generalized estimating equation (GEE) for total viable count (TVC), psychrophilic bacteria, total volatile base nitrogen (TVB-N), polyunsaturated fatty acids (PUFA), texture, taste, odor, and overall acceptance parameters. The results showed that by increasing the amount of chitosan, TVC, psychrophilic bacteria, and TVB-N decreased while PUFA, texture, taste, odor, and overall acceptance parameters increased.     

Conductive 3D structure nanofibrous scaffolds for spinal cord regeneration

The complex nature of spinal cord injuries has provided much inspiration for the design of novel biomaterials and scaffolds which are capable of stimulating neural tissue repair strategies. Recently, conductive polymers have gained much attention for improving the nerve regeneration. In our previous study, a three-dimensional (3D) structure with reliable performance was achieved for electrospun scaffolds. The main purpose in the current study is formation of electrical excitable 3D scaffolds by appending polyaniline (PANI) to biocompatible polymers. In this paper, an attempt was made to develop conductive nanofibrous scaffolds, which can simultaneously present both electrical and topographical cues to cells. By using a proper 3D structure, two kinds of conductive scaffolds are compared with a non-conductive scaffold. The 3D nanofibrous core-sheath scaffolds, which are conductive, were prepared with nanorough sheath and aligned core. Two different sheath polymers, including poly(lactic-co-glycolic acid) PLGA and PLGA/PANI, with identical PCL/PANI cores were fabricated. Nanofibers of PCL and PLGA blends with PANI have fiber diameters of 234±60.8 nm and 770±166.6 nm, and conductivity of 3.17×10^-5 S/cm and 4.29×10^-5 S/cm, respectively. The cell proliferation evaluation of nerve cells on these two conductive scaffolds and previous non-conductive scaffolds (PLGA) indicate that the first conductive scaffold (PCL/ PANI-PLGA) could be more effective for nerve tissue regeneration. Locomotor scores of grafted animals by developed scaffolds showed significant performance of non-conductive 3D scaffolds. Moreover, the animal studies indicated the ability of two new types of conductive scaffolds as spinal cord regeneration candidates.     

Investigating the reduction of sodium adsorption ratio from agricultural waste by rice husk in batch scale and physical model of drain envelop

Sodium adsorption ratio (SAR) is one of the water quality indexes that whose is important due to reuse or depletion to environment. Solutes in drain water can be controlled by adsorption, chemical or biological reaction, organic envelope of drainage. Rice husk is the common option of drainage envelops in paddy fields. In this study, the ability of reduction of SAR by rice husk was evaluated in batch scale and physical model of drain envelops. In the batch experiments, the adsorption of SAR parameters was investigated by adding 2 g of rice husk into a 100 ml of sodium chloride solution. The results indicated that rice husk absorbed calcium, magnesium and sodium, respectively. By increasing the temperature, contact time and pH, adsorption of calcium, magnesium and sodium was increased; however, the higher concentration of sodium in soil solution reduced the percentage of adsorption. In a more realistic state, physical models of subsurface drainage in the paddy fields were made. Drainage envelope treatments included of rice husk (H), combination of 20 and 60 % of husk with gravel (H₂₀G₈₀ and H₆₀G₄₀) and a pipe without envelope (NE). Due to higher drain discharge and more sodium removal (lower SAR in drain water), treatment H with the discharge of 16.2 ml/min and SAR of 1.27 (meq/l)^0.5 was better in comparison with other treatments.     

Retracted: Study of testis histology and sperm morphology in the Bunnei,Barbus sharpeyi (Gunther, 1874) induced by luteinizing hormone‐releasing hormone analogue and carp pituitary extract

This study was conducted to determine the effects of hormone treatment on testis structure in Barbus sharpeyi, as well as the morphology of sperm examined by scanning electronic microscopy (SEM). Male B. sharpeyi were divided into three groups (three fish per group) and injected with luteinizing hormone – releasing hormone analogue (LHRH–A₂) or carp pituitary extract (CPE). The first and second groups were treated with 10 μg kg^?1 LHRH–A₂ and metoclopramide (MET), and their testis were sampled pre‐ and Poststripping respectively. The third group received 2 mg kg^?1 CPE and were killed pre‐stripping. Based on the histological results obtained, the testicular connective tissue of the lumen was thicker, and seminal vesicles were of a lower volume, in fish injected with CPE in comparison to the other groups. After treatment with LHRH–A₂ and MET, not all spermatozoa within the testis were ejaculated, and only a small amount of sperm was obtained by abdominal stripping. The highest and lowest diameters of connective tissue within lobules were observed in fish receiving CPE and LHRH–A₂ treatments respectively. There was a significant difference (P < 0.05) in lobule space between the fish injected with the CPE and the fish injected with the LHRH–A₂ and MET. The SEM results revealed that the spermatozoa of B. sharpeyi were composed of a spherical to elliptical head, a cylindrical midpiece, and a lengthy flagellum. In conclusion, it was found that injection with LHRH–A₂ and MET improved the spermatogenic process in comparison to injection with CPE.