Suitability of tetrazolium test for Tamarindus indica L. seeds

Seed quality is routinely assessed by direct tests, e.g, the germination test, or indirect tests like the tetrazolium test, which has shoown to be promising in the determine viability and vigor, allowing the diagnosis of the main problems that may affect seed quality, such as mechanic damages, field deterioration and storage. In this respect, this study was conducted to develop a tetrazolium test protocol to evaluate the viability and vigor of Tamarindus indica L. seeds. Before exposing the seeds to the tetrazolium solution, seed preconditioning studies were carried out in which seven soaking times were tested. The soaking time that did not cause damage to the seed embryo and allowed the removal of the seed coat to expose the seed structures to the tetrazolium salt was selected. Then, an experiment was set up in a completely randomized design with a 2x3x3 factorial arrangement involving two seed lots, three soaking times in tetrazolium salt (6, 12 and 16 h) and three salt concentrations (0.075, 0.1 and 0.5%), totaling 18 treatments with four replicates of 25 seeds, evaluated at 40 ºC. For each treatment, the seeds were divided into three classes, namely, viable and vigorous embryos (class 1); viable embryos (class 2) and non-viable or dead embryos (class 3). For a comparison with the tetrazolium test results, the two seed lots were analyzed for water content, germination, emergence, first count, germination speed index, emergence speed index, growth and seedling dry weight. The viability and vigor of T. indica seeds can be evaluated after a soaking period of 48 h and immersion for 6h in tetrazolium salt at the concentration of 0.1%, at 40°C, with provides results similar to conventional seed viability tests. The tetrazolium test proved to be adequate to differentiate T. indica seed lots in terms of viability.

Extraction of chitin from edible crab shells of Callinectes sapidus and comparison with market purchased chitin

Chitin and its derived products have immense economic value due to their vital role in various biological activities as well as biomedical and industrial application. Insects, microorganism and crustaceans are the main supply of chitin but the crustaceans shell like shrimp, krill, lobsters and crabs are the main commercial sources. Chitin content of an individual varies depending on the structures possessing the polymer and the species. In this study edible crabs’ shells (Callinectes sapidus) were demineralized and deproteinized resulting in 13.8% (dry weight) chitin recovery from chitin wastes. FTIR and XRD analyses of the experimental crude as well as purified chitins revealed that both were much comparable to the commercially purchased controls. The acid pretreatment ceded 54g of colloidal chitin that resulted in 1080% of the crude chitin. The colloidal chitin was exploited for isolation of eighty five chitinolytic bacterial isolates from different sources. Zone of clearance was displayed by the thirty five isolates (41.17%) succeeding their growth at pH 7 on colloidal chitin agar medium. Maximum chitinolytic activity i.e. 301.55 U/ml was exhibited by isolate JF70 when cultivated in extracted chitin containing both carbon and nitrogen. The study showed wastes of blue crabs can be utilized for extraction of chitin and isolation of chitinolytic bacteria that can be used to degrade chitin waste, resolve environmental pollution as well as industrial purpose.

Efficacy of Purpureocillium lilacinum AUMC 10149 as biocontrol agent against root-knot nematode Meloidogyne incognita infecting tomato plant

Root-knot nematode Meloidogyne incognita is among the biotic factors which has greatly affected both the yield and the quality of the tomato crop. The egg parasitic nematode, Purpureocillium lilacinum (Pl) is considered as one of the most promising agents in controlling and overcoming this plant pathogen. The nematicidal effect of the native isolate Pl AUMC 10149 on second stage juvenile’s survival and egg hatching of M. incognita at different times of exposure was tested in vitro. The obtained data showed that Pl gave a maximum percentage of J2 mortality (97.6%) and egg hatching inhibition (79.8%) after 72 hours of exposure. The potentiality of Pl as well as Bio-Nematon to control M. incognita infecting tomato was conducted using different times of application in vivo. Nine treatments with five replicates were used for such bioagents compared with the nematicide Oxamyl. Each seedling was inoculated with 1000 J2s of nematode/pot and 10 mL of Pl (1x1010 CFU/mL) or Bio-Nematon spore suspension (1x108 CFU/mL) 10mL/pot. The results indicated that the most effective treatments in reducing nematode population, number of galls and egg masses of M. incognita in plant roots was performed with treatment by Pl pre-planting and post-infection with Pl (Rf 1.9) giving a significant enhancement in plant length (64.9%), fresh weight (72.52%) and shoot dry weight (163.41%) without negatively impacting environment. Therefore, the present study confirmed that using P. lilacinum AUMC 10149 can be used as a practical supplement to environmentally friendly disease management of root-knot nematodes in Egypt.

Fish Anchor Dived – Confirmation Through Field Tests in the Swan River

This paper reports the results from field tests on a 1/15th scale recently developed fish anchor. The tests were conducted at three locations in the Swan River, Perth. Two series of tests were performed from the Burswood and Maylands jetties with water depths between 1.1 and 1.9 m. The final series of tests were undertaken in deeper waters of 2.6 m from a barge. The riverbed at the Burswood Jetty and barge test location consisted of soft clay, and that at the Maylands Jetty comprised sandy silt. The tip embedment depths of the scaled fish anchor, with dry weight of 0.304 kN and impact velocity of 5.89∼9.55 m/s, in soft clay were 1.17∼2.40 times the anchor length. For similar impact velocities, the tip embedment depths in sandy silt were 30 ∼ 60% shallower than those in soft clay. By comparing the field test data in clay, the fish anchor achieved normalised embedment depths similar to those of the torpedo and OMNI-Max anchors under half or less impact velocity. Most importantly, the field tests confirmed the diving behaviour of the fish anchor under loading with mudline inclination of 20° and 25°, with the second peak dictated the capacity. The ultimate capacity was 5∼7 times the anchor submerged weight in water.

Nest refuse of Acromyrmex balzani (Hymenoptera: Formicidae) increases the plant vigor in Turnera subulata (Turneraceae)

Some studies report the positive effect of organic residues from ant nests on soil properties and on the structure of the adjacent plant community in field experiments, but there is a gap about the effect on individual species. The purpose of the present study was to compare the soil nutrient content and the development of Turnera subulata Smith, an ornamental species, in the presence of the nest refuse (basically composed of fragments of grass leaves and the symbiotic fungus) produced by the leaf-cutting ant Acromyrmex balzani (Emery, 1890) or in control soil through a greenhouse pot experiment. The experiment was carried out with two treatments: control soil and soil with 25% of nest refuse. The plants were kept in 1L pots for 90 days. We evaluated the parameters: plant height, stem diameter, root length, number of leaves, dry weight of the root, dry and fresh aboveground biomass. Additionally, the relative chlorophyll content and leaf nutrients were used as nutritional parameters. As a result, plants that grew in the soil with nest refuse showed significant higher values of all parameters evaluated when compared to the control treatment (p < 0.001). We conclude that this biofertilizer contributed to the production of more vigorous plants, being able to act on the local dynamics of nutrients in the ecosystems where A. balzani occurs. As it is relatively abundant and easy to collect, the refuse of A. balzani has the potential to be used as an alternative substrate in the production of shortlife cycle plants.

A Library of Aspergillus niger Chassis Strains for Morphology Engineering Connects Strain Fitness and Filamentous Growth With Submerged Macromorphology

Submerged fermentation using filamentous fungal cell factories is used to produce a diverse portfolio of useful molecules, including food, medicines, enzymes, and platform chemicals. Depending on strain background and abiotic culture conditions, different macromorphologies are formed during fermentation, ranging from dispersed hyphal fragments to approximately spherical pellets several millimetres in diameter. These macromorphologies are known to have a critical impact on product titres and rheological performance of the bioreactor. Pilot productivity screens in different macromorphological contexts is technically challenging, time consuming, and thus a significant limitation to achieving maximum product titres. To address this bottleneck, we developed a library of conditional expression mutants in the organic, protein, and secondary metabolite cell factory Aspergillus niger. Thirteen morphology-associated genes transcribed during fermentation were placed via CRISPR-Cas9 under control of a synthetic Tet-on gene switch. Quantitative analysis of submerged growth reveals that these strains have distinct and titratable macromorphologies for use as chassis during strain engineering programs. We also used this library as a tool to quantify how pellet formation is connected with strain fitness and filamentous growth. Using multiple linear regression modelling, we predict that pellet formation is dependent largely on strain fitness, whereas pellet Euclidian parameters depend on fitness and hyphal branching. Finally, we have shown that conditional expression of the putative kinase encoding gene pkh2 can decouple fitness, dry weight, pellet macromorphology, and culture heterogeneity. We hypothesize that further analysis of this gene product and the cell wall integrity pathway in which it is embedded will enable more precise engineering of A. niger macromorphology in future.

Fast-Tracking Isolation, Identification and Characterization of New Microalgae for Nutraceutical and Feed Applications

Microalgae contain high-value biochemical compounds including fatty acids (FA), protein and carotenoids, and are promising bioresources to enhance nutrition of food and animal feed. Important requirements for commercial strains are rapid growth and high productivities of desirable compounds. As these traits are believed to be found in aquatic environments with fluctuating conditions, we collected microalgae from marine and freshwater environments that are subjected to eutrophication and/or tidal fluctuations. Using this directed approach, 40 monoalgal cultures were isolated and 25 identified through 18S rDNA sequencing and morphological characterization. Based on their high growth rates (0.28–0.60 day−1) and biomass productivities (0.25–0.44 g L−1day−1) in commercial fertilizer under standardized conditions, six new strains were selected. Scenedesmus sp. GW63 produced quality FA-rich biomass with high omega-3 polyunsaturated FA (28.5% of total FA (TFA)) contents, especially α-linolenic acid (ALA; 20.0% of TFA) with a very low n-6/n-3 ratio (0.4), and high FA productivity (32.6 mg L−1 day−1). A high protein productivity (34.5 mg L−1 day−1) made Desmodesmus sp. UQL1_26 (33.4% of dry weight (DW)) attractive as potential protein-rich feed and nutrition supplement. Monoraphidium convolutum GW5 displayed valuable carotenoid production (0.8% DW) with high carotenoid accumulation capability (0.8 mg L−1 day−1). This research provides a pathway for fast-tracking the selection of high-performing local microalgae from different environments for nutraceuticals, functional foods and animal feed applications.

Comparative Characterization and Identification of Poly-3-Hydroxybutyrate Producing Bacteria with Subsequent Optimization of Polymer Yield

In this work, the strains Bacillus megaterium BM 1, Azotobacter chrocococcumAz 3, Bacillus araybhattay RA 5 were used as an effective producer of poly-3-hydroxybutyrate P(3HB). The purpose of the study was to isolate and obtain an effective producer of P(3HB) isolated from regional chestnut soils of northern Kazakhstan. This study demonstrates the possibility of combining the protective system of cells to physical stress as a way to optimize the synthesis of PHA by strains. Molecular identification of strains and amplification of the phbC gene, transmission electron microscope (TEM), extracted and dried PHB were subjected to Fourier infrared transmission spectroscopy (FTIR). The melting point of the isolated P(3HB) was determined. The optimal concentration of bean broth for the synthesis of P(3HB) for the modified type of Bacillus megaterium RAZ 3 was 20 g/L, at which the dry weight of cells was 25.7 g/L−1 and P(3HB) yield of 13.83 g/L−1, while the percentage yield of P(3HB) was 53.75%. The FTIR spectra of the extracted polymer showed noticeable peaks at long wavelengths. Based on a proof of concept, this study demonstrates encouraging results.

Effect of Nitrogen Concentration on the Alkalophilic Microalga Nitzschia sp. NW129-a Promising Feedstock for the Integrated Production of Lipids and Fucoxanthin in Biorefinery

Microalgae are considered promising resources for producing a variety of high-value-added products, especially for lipids and pigments. Alkalophilic microalgae have more advantages than other microalgae when cultured outdoors on a large scale. The present study investigated the comprehensive effects of different nitrogen concentrations on fucoxanthin (Fx), lipids accumulation and the fatty acid profile of the alkaliphilic microalgae Nitzschia sp. NW129 to evaluate the potential for simultaneous production of Fx and biofuels. Fx and Lipids amassed in a coordinated growth-dependent manner in response to various concentrations, reaching 18.18 mg g–1 and 40.67% dry weight (DW), respectively. The biomass of Nitzschia sp. NW129 was 0.58 ± 0.02 g L–1 in the medium at the concentration of 117.65 mM. The highest productivities of Fx (1.44 mg L–1 d–1) and lipid (19.95 ± 1.29 mg L–1 d–1) were obtained concurrently at this concentration. Furthermore, the fatty acid methyl esters revealed excellent biofuel properties with an appropriate value of the degree unsaturation (49.97), cetane number (62.72), and cold filter plugging point (2.37), which met the European standards for biofuel production (EN14214). These results provided a reliable strategy for further industrialization and comprehensive production of biofuel and Fx by using the alkaliphilic microalgal Nitzschia sp. NW129.