Establishing bacterial infectivity models in striped Catfish Pangasianodon hypophthalmus (Sauvage) with Edwardsiella ictaluri

A bacterial infectivity challenge model of Edwardsiella ictaluri in striped catfish was developed. All experiments were conducted using a bacterial isolate of E. ictaluri that had been recovered during a natural outbreak of bacillary necrosis of Pangasianodon (BNP) in farmed striped catfish Pangasianodon hypophthalmus in Vietnam. Time of immersion in 10⁷ CFU.ml⁻¹ had a significant effect on mortality. The immersion bacterial dose of 10⁷ CFU/ml for 30 s resulted in a cumulative percentage mortality of 63%. Three to four days post-bacterial challenge, fish showed gross clinical signs of natural BNP and E. ictaluri was recovered and identified from these fish. Moreover, a cohabitation challenge was evaluated as an alternative challenge method, although the mortalities among the infected fish were lower at around 15%–40%. This study confirmed the horizontal transmission of E. ictaluri in striped catfish and elucidated that cohabitation challenge could be used in reproducing the disease under controlled conditions.     

Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Genetic variation in root development responses to salt stresses of quinoa

Soil salinity has become a serious environmental abiotic stress limiting crop productivity and quality. The root system is the first organ sensing the changes in salinity. Root development under elevated salinity is therefore an important indicator for saline tolerance in plants. Previous studies focused on varietal differences in morphological traits of quinoa under saline stresses; however, variation in root development responses to salinity remains largely unknown. To understand the genetic variation in root development responses to salt stress of quinoa, we conducted a preliminary screening for salinity response at two salinity levels of a diverse set of 52 quinoa genotypes and microsatellite markers were used to link molecular variation to that in root development responses to salt stresses of represented genotypes. The frequency distribution of saline tolerance index showed continuous variation in the quinoa collection. Cluster analysis of salinity responses divided the 52 quinoa genotypes into six major groups. Based on these results, six genotypes representative of groups I to VI including Black quinoa, 2-Want, Atlas, Riobamba, NL-6 and Sayaña, respectively, were selected to evaluate root development under four saline stress levels: 0, 100, 200 and 300 mM NaCl. Contrasts in root development responses to saline stress levels were observed in the six genotypes. At 100 mM NaCl, significant differences were not observed in root length development (RLD) and root surface development (RSAD) of most genotypes except Black quinoa; a significant reduction was observed in this genotype as compared to controls. At 200 mM NaCl, significant reduction was detected in RLD and RSAD in all genotypes showing this as the best concentration to discriminate among genotypes. The strongest inhibition of root development was found for all genotypes at 300 mM NaCl as compared to lower saline levels. Among genotypes, Atlas of group III shows as a saline-tolerant genotype confirming previous reports. Variation in root responses to salinity stresses is also discussed in relation to climate conditions of origins of the genotypes and reveal interesting guidelines for further studies exploring the mechanisms behind this aspect of saline adaptation.     

Naringin-loaded Poly(ε-caprolactone)/Gelatin Electrospun Mat as a Potential Wound Dressing: In vitro and In vivo Evaluation

In the present study, naringin, a flavonoid isolated from the grape and citrus fruit species, was incorporated with poly(ε-caprolactone)/gelatin composite mats in order to develop a potential wound dressing. The composite mats were prepared by electrospinning of poly(ε-caprolactone)/gelatin (1:1 (w/w)) solution incorporated with 1.50 %, 3 % and 6 % (w/w) of naringin. The electrospun mats were evaluated regarding their morphology, contact angle, water-uptake capacity, water vapor transmission rate, tensile properties, drug release, cellular response and in vivo wound healing activity. The study showed that after 2 weeks, the full-thickness excisional wounds of Wistar rats treated with the naringin-loaded dressings achieved a wound closure of higher than 94 % and the dressing containing 6 % (w/w) naringin had almost 100 % wound closure. The sterile gauze, as the control group, showed nearly 86 % of wound closure after this period of time. Our results provided evidence that supports the possible applicability of naringin-loaded wound dressing for successful wound treatment.     

Swim bladder inflation as a possible cause of saddleback-like syndrome malformation in hatchery-reared red spotted grouper <Emphasis Type="Italic">Epinephelus akaara</Emphasis>

Saddleback-like syndrome is one of the most frequently observed malformations in hatchery-reared red spotted grouper Epinephelus akaara. Saddleback-like syndrome is characterized by a V-shaped dorsal-profile depression behind the head region accompanied by deformities or misalignments of the first to fourth neural spines on the vertebral column and the first to fourth pterygiophores of the dorsal fin. Abnormal swim bladder inflation in several finfish species leads to malformations, particularly of the vertebral column. The present study examined the influence of swim bladder inflation on saddleback-like syndrome in hatchery-reared red spotted grouper. The incidence of saddleback-like syndrome increased as swim bladder inflation rate increased. Additionally, the incidence of saddleback-like syndrome among juveniles with inflated swim bladders was significantly higher than that among juveniles with uninflated swim bladders. The larval swim bladder is located just below the first to fifth vertebrae and the first to fifth dorsal spines, where saddleback-like syndrome occurs. Our results suggest that an inflated swim bladder increases the risk of saddleback-like syndrome in hatchery-reared red spotted grouper.     

Cylindrospermopsin effects on cell viability and redox milieu of Neotropical fish <Emphasis Type="Italic">Hoplias malabaricus</Emphasis> hepatocytes

Cylindrospermopsin (CYN) is a cyanotoxin that is cytotoxic to a wide variety of cells, particularly to the hepatocytes. In this study, the toxic effects of purified CYN were investigated in primary cultured hepatocytes of Neotropical fish Hoplias malabaricus. After isolation, attachment, and recovery for 72 h, the cells were exposed for 72 h to 0, 0.1, 1.0, 10, and 100 μg l^?1 of CYN. Then, cell viability and a set of oxidative stress biomarker responses were determined. Catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione S-transferase activities were not affected by exposure to CYN. Concentration-dependent decrease of glutathione reductase activity occurred for most CYN-exposed groups, whereas non-protein thiol content increased only for the highest CYN concentration. Lipid peroxidation, protein carbonylation, and DNA damage levels were not altered, but reactive oxygen species levels increased in the cells exposed to the highest concentration of CYN. Cell viability decreased in all the groups exposed to CYN. Thus, CYN may cause a slight change in redox balance, but it is not the main cause of cell death in H. malabaricus hepatocytes.     

Androgen-induced pseudo-hermaphroditic phenotypes in female <Emphasis Type="Italic">Brevimyrus niger</Emphasis> Günther 1866 (Teleostei,Mormyridae)

This paper explores the plasticity of sexually dimorphic characters in subadult female Brevimyrus niger, an African weakly electric mormyrid species. Thirty-five fish were exposed in a staggered fashion (five fish a week) to aromatizable 17α-methyltestosterone over a period of 7 weeks; 18 fish served as untreated controls. 17α-MT induced precocious vitellogenesis that mirrored the natural maturational process during seasonal ovarian recrudescence. At the same time, 17α-MT exposure resulted in complete masculinization of the females’ anal fin support structure normally observed during rainy season in adult males. We discuss possible hormonal mechanisms acting along the brain-pituitary-gonad axis that would explain the occurrence of precocious vitellogenesis and the male-typical transformation of the female’s anal fin ray bases. Our findings are relevant to commercial aquaculture as the use of 17α-MT in fish hatcheries can pose serious environmental issues.     

Free-Living Plant-Parasitic Nematodes do not Affect the Efficiency of Seed Tuber Fungicide Treatment against <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Stem canker on germinating potato sprouts is often caused by seed-borne inoculum of the fungus Rhizoctonia solani. However, high amounts of free-living plant-parasitic nematodes have been found in field patches of potato plants with stem canker. Fungicide treatment of the seed tubers can be used to avoid stem canker caused by seed-borne inoculum but it is unknown if nematodes can affect this. To investigate whether free-living plant-parasitic nematodes, the root-lesion nematode Pratylenchus penetrans or a combination of several plant-parasitic nematode genera in a full nematode community, may have a negative effect on the fungicide seed treatment, a pot experiment with seed tubers inoculated with R. solani, half of which were treated with fungicides, was performed. The seed-borne inoculum caused severe damage to the plants, while no fungal damages were observed on the fungicide treated plants. This shows that the nematodes did not affect the fungicide treatment. The probability of black scurf decreased in treatments with a full nematode community, which may be due to the action of fungal-feeding nematodes.