M4 Medium Research Articles

Toxicity Evaluation of Copper, Nickel, and Mixture on Daphnia Magna

This study conducted acute (48 hours) and chronic (21 days) toxicity tests of nickel (Ni2+), copper (Cu2+) ions, and a mixture of the Ni2+ - Cu2+ ions on Daphnia magna (D. magna) under M4 medium. The acute test results showed that the toxicity of Cu2+ was about 15 times higher than Ni2+, demonstrated by the 50% effect concentration (EC50) value of 185.2 μg.L-1 and 2706.97 μg.L-1. In addition, the results also illustrated that the mixture of the Cu2+ – Ni2+ ions was more toxic to D. magna than a single metal with an EC50 value of 175.22 μg.L-1. Acute toxicity tests showed that the metal affected the viability of D. Magna, while the organism maturation and reproduction were affected under chronic exposure. Chronic test results showed that the toxicity of Cu2+ was higher than that of Ni2+ with EC50 values of 9.06 μg.L-1 and 162.12 μg.L-1; noticeably, the toxicity of the mixture of the two metals is higher than that of the single metal. In particular, Ni2+ at a concentration of 100 μg.L-1 stimulated the maturation, survival, and reproduction of D. magna. but at higher concentrations (> 100 μgNi.L-1), Ni2+ would bind to Cu2+, which exerted a more substantial effect on the test organism. This study initially evaluated the toxicity of Cu2+ and Ni2+ on microcrustaceans D. magna, which is the premise for further studies on the genotoxicity of heavy metals on microcrustaceans in general and D. magna in particular to ensure the quality of the ecosystem.

Production, Characterization, Kinetics, and Thermodynamics Analysis of Amyloglucosidase from Fungal Consortium.

The current study aimed to produce an amyloglucosidase enzyme from the fungal consortium. The best amylolytic fungal consortia were identified as Alternaria alternata and Aspergillus niger through the 18S rDNA technique. Fermentation kinetics and various nutritional and cultural parameters were analyzed. Maximum production was obtained in M4 media, pH 5.5, 30°C, and 4mL inoculum at 150rpm after 72h of incubation. Along with that, sodium nitrate at 2.5%, maltose, beef extract 1%, zinc sulfate (0.1%), and Tween 80 (0.1%) supported the maximum amyloglucosidase production. Amyloglucosidase was partially purified up to 1.6 purification fold with a specific activity of 1.84 Umg-1 in a stepwise manner by ammonium sulfate purification, dialysis, and ion exchange chromatography. The AMG enzyme also revealed maximum activity at 50°C with 5.0 pH. Upon the kinetic analysis, the specific yield coefficient Yp/x and volumetric rates Qp and Qx were also found to be significant in the above optimized conditions. The Km value 0.33mgmL-1 and Vmax 26.31 U mL-1 were obtained at 1% soluble starch substrate. Thermodynamic parameters for soluble starch hydrolysis were as follows: ΔH = 48.78kJmol-1, (Ea) = - 46.0kJmol-1, and ΔS = - 43.10Jmol-1K-1. This finding indicates the indigenously isolated fungal consortium can be the best candidate for industrial applications.

Control of Microcystis aeruginosa by Daphnia: Experimental evidence and identification of involved infochemicals.

Infochemicals refer to chemicals responsible for information exchange between organisms. We evaluated the effects of Daphnia magna and Daphnia galeata infochemicals on Microcystis aeruginosa for 15d. The Daphnia infochemicals were obtained from spent medium after culturing Daphnia in Elendt M4 medium for 48 h. Both Daphnia infochemicals significantly increased (p < 0.05) the intracellular reactive oxygen species level and microcystin-LR concentration in M. aeruginosa. This cellular effect increased colony formation of M. aeruginosa, thereby inhibiting the growth of M. aeruginosa. D. galeata infochemicals provoked significantly greater (p < 0.05) adverse effects on M. aeruginosa than those of D. magna infochemicals, which were further exaggerated by pre-exposure of Daphnia to M. aeruginosa. This result seems to be related to the different compositions and concentrations of Daphnia infochemicals. Several Daphnia infochemicals, such as methyl ferulate, cyclohexanone, 3, 5-dimethyl, hexanedioic acid, and bis(2-ethylhexyl) ester, showed a high correlation with M. aeruginosa cell concentration (|r | >0.6), suggesting that they may play a key role in controlling harmful cyanobacteria. Additionally, pre-exposure of D. magna and D. galeata to M. aeruginosa produced oleic acid, methyl ester, and n-hexadecanoic acid, with a highly correlation with M. aeruginosa cell concentration (|r | >0.6). p-tolyl acetate and linoleic acid were detected only in the pre-exposed D. galeata infochemicals. These findings suggest that some of Daphnia infochemicals identified in this study can be a promising tool to control M. aeruginosa growth. However, further studies are required to verify the specific actions of these infochemicals against cyanobacteria.

Factors Affecting the Micropropagation of Sapindus trifoliatus from Nodal Explants of Mature Tree

Background:: Explant recalcitrance is one of the major challenges during the tissue culture of a tree species, especially when explants are derived from mature plants. It is mainly as-sociated with the genotypes/species, source and physiological status of explants, seasonal variations, and competency/incompetency of explants towards PGRs for in vitro manipulations. There-fore, to optimize the appropriate conditions and minimize explant recalcitrance, it is necessary to elucidate the different factors influencing the tissue culture of a tree species. Objective:: Several factors influencing in vitro shoot induction from nodal explants derived from mature plants of Sapindus trifoliatus were investigated. Methods:: Nodal segment obtained from mature plants was used as an explant for shoot regeneration. The factors assessed in this study included the types and concentrations of plant growth regulators, nutrient composition of MS (Murashige and Skoog) medium, sucrose concentrations, the position of nodes on their respective branches, seasonal variations, and successive transfers of mother explant. Results:: The performance of nodal explants was better on modified MS medium, i.e., M4 medium (half-strength of MS major salts and full-strengths of MS minor salts, Fe-EDTA and MS organic nutrients each) than the full-strength MS medium. The addition of BAP in the medium resulted in a higher shoot induction rate than other cytokinins i.e., Kin, Zeatin, TDZ. M4 medium supplemented with 3.0 mg l-1 BAP and 2% sucrose was optimum for shoot proliferation. However, elongation of shoots was observed only when induced shoots, along with the mother explant, were subcultured on a medium containing 1.0 mg l-1 BAP. The shoot multiplication was positively affected by the repeated transfer of the mother explant for different passages on the multiplica-tion medium. Mother explant transferred to multiplication medium (0.8% agar-solidified M4 medium + 2% sucrose + 1.0 mg l-1 BAP) proliferated maximum shoots after fourth passages. Regenerated shoots were rooted in vitro and further successfully acclimatized in field conditions. Conclusion:: The in vitro regeneration system developed from an explant obtained from mature trees can be successfully exploited for mass multiplication of Sapindus trifoliatus, a medicinally and commercially important tree.

Effectiveness of media B5 (Gamborg) for multiplication of Sengon (Paraserianthes falcataria L. Nielsen) in vitro

Paraserianthes falcataria (L.) Nielsen, also known as sengon, is one of the most important and adaptable pioneer species. The Sengon plant is one of the quickest-growing tropical plants. The objective of this study was to identify the optimal medium for sengon multiplication with the addition of ZPT, cytokinin, BAP, and kinetin, either singly or in combination. The research design employed was an entirely random design (CRD). Media M2 (B5 + BAP 0.1), which was the medium with the highest number of shoots, was determined to be the optimal medium for sengon plantlets in the study. At 1 MST, the fastest medium for leaf emergence was (B5 + BAP 0.2) media. The poisson test analysis shows that, in comparison to other media, the M2 medium (B5 + BAP 0.1) has a very significant impact on the quantity of leaves. The medium with the highest average height of sengon plantlets was M1 (B5 Control), with 1.73 cm. The highest percentage of dead plantlets on M4 medium was 33%, while on M9 medium it was 17%, due to browning leading to cell death. Only media M4 (BAP 0.2), M5 (Kinetin 0.2), and M9 (BAP (0.2) + Kin (0.2) contained plantlets contaminated by bacteria at a concentration of up to 17%, which inhibited plantlet growth.

Development of an in vitro regeneration system from immature inflorescences and CRISPR/Cas9-mediated gene editing in sudangrass

BackgroundSudangrass (Sorghum sudanense) is a major biomass producer for livestock feed and biofuel in many countries. It has a wide range of adaptations for growing on marginal lands under biotic and abiotic stresses. The immature inflorescence is an explant with high embryogenic competence and is frequently used to regenerate different sorghum cultivars. Caffeic acid O-methyl transferase (COMT) is a key enzyme in the lignin biosynthesis pathway, which limits ruminant digestion of forage cell walls and is a crucial barrier in the conversion of plant biomass to bioethanol. Genome editing by CRISPR/Cas9-mediated mutagenesis without a transgenic footprint will accelerate the improvement and facilitate regulatory approval and commercialization of biotech crops. Methods and resultsWe report the overcome of the recalcitrance in sudangrass transformation and regeneration in order to use genome editing technique. Hence, an efficient regeneration system has been established to induce somatic embryogenesis from the immature inflorescence of two sudangrass cultivars on four MS-based media supplemented with different components. Our results indicate an interaction between genotype and medium composition. The combination of Giza-1 cultivar and M4 medium produces the maximum frequency of embryogenic calli of 80% and subsequent regeneration efficiency of 22.6%. Precise mutagenesis of the COMT gene is executed using the CRISPR/Cas9 system with the potential to reduce lignin content and enhance forage and biomass quality in sudangrass. ConclusionA reliable regeneration and transformation system has been established for sudangrass using immature inflorescence, and the CRISPR/Cas9 system has demonstrated a promising technology for genome editing. The outcomes of this research will pave the road for further improvement of various sorghum genotypes to meet the global demand for food, feed, and biofuels, achieving sustainable development goals (SDGs).

Fe2O3-Ag2O/TiO2 Nanocatalyst-Assisted LC-MS/MS-Based Detoxification of Pesticide Residues in Daphnia magna and Algae Mediums

In this work, a simple sensitive validated liquid chromatography mass spectroscopy (LC-MS/MS) analytical method was developed for the determination of Spirodiclofen residues in different aquatic toxic media. The toxic media were those that provide nutrients and help with the growth of different aquatic organisms for their survival and multiplication. The different media were the M4 medium for Daphnia magna and The Organization for Economic Cooperation and Development (OECD TG 201) medium for alga. Fe2O3-Ag2O/TiO2 nanocomposites were prepared by using a precipitation method, which was used as a photo-catalyst for the removal of Spirodiclofen pesticide from aquatic media. The experiment was performed under direct sunlight at a single fortification level (1.0 µg/mL) in M4 and OECD TG 201 media. The optimum catalyst concentration for the complete degradation was found to be 10 mg/L under sunlight. Spirodiclofen residues in water were determined by LC-MS/MS, and the rate constant DT50 (half-life) values were calculated from the obtained data. The results showed that with Fe2O3-Ag2O/TiO2 nanocatalyst, the DT50 (half-life) value was found to be approximately 8 h. These results revealed that iron-oxide- and silver-oxide-incorporated TiO2 nanocomposites were excellent photocatalysts when compared with TiO2, Fe2O3-TiO2, and Ag2O-TiO2 for the decontamination of pesticide residues in aquatic media samples.

Maternal Effect of Polyethylene Microplastic Fragments Containing Benzophenone-3 in Different Ages and Broods of Daphnia Magna

The maternal effect of microplastics (MPs) toxicity is likely influenced by age and brood of test species. This study investigated the maternal effect of polyethylene MP fragments (18.23 ± 8.02μm) with benzophenone-3 (BP-3; 2.89 ± 0.20% w/w) on chronic toxicity to Daphnia magna over two generations. Neonate (< 24h old) and adult (5 d old) daphnids in the F0 generation were exposed until 21 d old, then first and third brood neonates in the F1 generation were recovered in clean M4 medium for 21 d. Higher chronic toxicity and maternal effect of MP/BP-3 fragments were observed in the adult group compared with the neonate group, reducing growth and reproduction in both F0 and F1 generations. First brood neonates in the F1 generation showed a higher maternal effect of MP/BP-3 fragments than third brood ones, resulting in enhanced growth and reproduction relative to the control. This study provided insights into the ecological risk of MPs containing plastic additives in the natural environment.

Elimination of coexisting canna yellow mottle virus, bean yellow mosaic virus and cucumber mosaic virus from Canna generalis cv. black knight through in vitro chemotherapy of rhizome explants.

During our previous study, the mixed infection of canna yellow mottle virus (CaYMV), bean yellow mosaic virus (BYMV), and cucumber mosaic virus (CMV) was identified in a Black Knight cultivar of canna exhibiting severe yellow streak and mottling symptoms. Before the development of the virus-free plants, the ability of callogenesis and organogenesis from the ovary, stalk, and rhizome explants was tested on different concentrations and combinations of TDZ, NAA, BAP, and Ads growth regulators. The performance of rhizome explants was above all the explant types and 33.33 ± 1.67 rhizomes (out of 50 placed) showed callus development on ME medium (MS supplemented with 0.8mg/L TDZ and 0.25mg/L NAA) and further on a refined M4 medium (MS supplemented with 4.0mg/L BAP, 1.0mg/L NAA and 50mg/L Ads) produced 4.06 ± 0.16 shoots per explant. The development of virus-free plants was attempted by in vitro chemotherapy using ribavirin. Not only in callogenesis and shoot development but also in the ribavirin treatments, rhizomes developed about 3.78 ± 0.68 shoots per explant on 40mg/L ribavirin in the ME medium. These optimizations suggested that ME medium for callogenesis, M4 medium for shoot development and the treatment of 40mg/L ribavirin for 30days at M4 medium was effective. The elimination of coinfection of all three viruses from rhizome explants of 0.5 cm2 of the Black Knight cultivar was attempted. Consequently, a total of 53.33% of plants free from all three viruses (48 out of the 90 plants developed) were obtained when screened by RT-PCR and PCR for their absence.

Seedling Growth and Quality of Avicennia marina (Forssk.) Vierh. under Growth Media Composition and Controlled Salinity in an Ex Situ Nursery

Avicennia marina (Forssk.) Vierh. is an important mangrove species that inhabits the outermost zone of mangrove forests, but it has been shown to have a poor ability to regenerate due to its low seedling quality. We conducted a study to evaluate the specific growth requirements of A. marina, i.e., medium and salinity level. Germinated seeds were transplanted to pots filled with media, i.e., silt loam (M1), loam (M2), sandy loam (M3), or sand (M4), with various salinity levels 5 (S1), 5–15 (S2), 15–25 (S3), or 25–35 ppt (S4). Survival rate, growth, biomass partition, and seedling quality were observed for 14 weeks after transplanting the seeds. The highest rate of seedling survival was found in the S2 condition, and higher concentrations of salinity lowered the survival rates. The S1 treatment promoted the initial 8 week growth of the seedlings. Growth medium had no significant effect, except on the survival rates grown in M4. Growth medium composition had no distinct effect on seedling growth. The S2 and S3 treatments induced better growth (in terms of shoot height and root length) and resulted in high-quality (i.e., Dickson quality index) seedlings in any type of medium. The S3 treatment increased the seedling quality in M1 and M4, whereas the S4 treatment only benefited seedlings in the M4 medium. According to the results, a specific range of salinity (5–15 ppt) with circulated water in any type of medium is recommended for the establishment of an ex situ nursery for the propagation of A. marina, in contrast to the general range of salinity (4–35 ppt) stated in previous references.

FIELD TEST OF THE INDIGENOUS MICROBES FOR OIL RECOVERY, LEDOK FIELD, CENTRAL JAVA

After selecting several old wells in Cepu, then well LDK-132 was chosen for implementing MEOR technology using "huff and puff" method. For this purpose, fluids samples (water and oil) from Ledok Formation were taken at the wellhead. No core of LDK-132 was available, only cores from LDK-209 and LDK-P1 were found. Core plugs could not be made from the former due to compacted and tightly conditions with a very low permeability of only 0.458 md. The latter with the permeability of 140 md met the requirements of microbial core flooding (MCF) tests, A standard quartz dominated core from Clashach Scotland with permeability almost 800 md was also used to compare with the native core's results. The MCF tests were conducted at the reservoir conditions for both types of core. The native core gave a recovery factor of oil production of 12.58%, while the standard core yielded a higher recovery fac- tor of 21.22% of Sor. Based on these results, the MEOR implementation was conducted on July 8, 1999 by injecting 135 barrels of mixtures consist- ing of formation water, microbes enriched with KKL-11 (Koleksi Kultur Lemigas or Lemigas Cultures Collection) and M4 medium through the annulus of LDK-132. The result showed an increase of average oil production rate, from 3.46 bopd, 6 days before the injection, to 24.85 bopd, 6 days after the injection.

The Alternative Media Supporting the Protocorm and Plantlet Growth of the Indonesian Black Orchid (Coelogyne pandurata Lindl.) Grown In Vitro

&lt;p&gt;Due to the high cost of the most often used basic media in tissue culture, it is vital to identify more affordable alternatives. This research aimed to determine the best alternative culture media for the protocorm and plantlet growth of &lt;em&gt;Coelogyne pandurata&lt;/em&gt; Lindl. It employed a completely randomized design, eight treatments and four replications. The treatments referred to the different types media and consisted of M1 = New Phalaenopsis (NP) medium, M2 = a medium made from foliar fertilizer (FFM), M3 = NP + 2 cc L&lt;sup&gt;-1&lt;/sup&gt; AB mix solution (a media for hydroponics), M4 = FFM + 2 cc L&lt;sup&gt;-1&lt;/sup&gt; AB mix solution, M5 = NP + 50 cc L&lt;sup&gt;-1&lt;/sup&gt; of tomato extract, M6 = NP + 50 cc L&lt;sup&gt;-1&lt;/sup&gt; of bean sprout extract, M7 = FFM + 50 cc L&lt;sup&gt;-1&lt;/sup&gt; of tomato extract and M8 = FFM + 50 cc L&lt;sup&gt;-1&lt;/sup&gt; of bean sprout extract. The M4 medium exhibited the best results in terms of average leaves count (4.80), average shoot length (2.68 cm), average root length (4.35 cm), the average fresh weight per plantlet (214.5 mg) and dry weight of plantlets (73.1 mg). The average number of roots per plantlet was 4.25, acquired using the less expensive M8 treatment, which also produces a negligible number of leaves (4.50). In conclusion, the M4 medium is the most appropriate medium for growing protocorm and plantlet of &lt;em&gt;C. pandurata&lt;/em&gt;. The experiment also found that the FFM basic medium combined with 50 cc L&lt;sup&gt;-1&lt;/sup&gt; of bean sprout extract can be used as another cheaper alternative for growing protocorms of &lt;em&gt;C. pandurata&lt;/em&gt;.&lt;/p&gt;

Ecotoxicological assessment of amoxicillin trihydrate: Stability, solubility, and acute toxicity for Oreochromis niloticus, Lemna minor, and Daphnia magna

Among the different pharmaceuticals present in water as micro-contaminants, antibiotics are considered an important risk group for the aquatic ecosystem, since their increasing use in human and veterinary medicine could affect the diversity of microorganism communities. This study aims to classify the acute toxicity of amoxicillin trihydrate for organisms of different trophic levels of aquatic biological chains, represented by Oreochromis niloticus (fish), Lemna minor (macrophyte) and Daphnia magna (microcrustacean). For this purpose, amoxicillin trihydrate degradability in water as well as changes in physicochemical characteristics are focused herein. Initial tests were performed to validate the assay, including drug dissolution in M4 medium, drug degradation in various culture media, sensitivity of experimental organisms as well as preliminary drug intoxication test. Upon validation tests, 2.6 g L−1 amoxicillin was determined as the concentration to evaluate the toxicity factor. Static (48 h) and semi-static (168 h) assays were performed. Therefore, amoxicillin trihydrate did not have an acute toxic effect at the concentration of 2.6 g L−1 under the experimental conditions applied to the species Lemna minor, Daphnia magna and Oreochromis niloticus. demonstrating the very low acute toxicity to aquatic organisms at different trophic levels.

Gamma cyclodextrin glycosyltransferase from evansella caseinilytica: production, characterization and product specificity.

Alkalohalophilic Evansella caseinilytica produced an extracellular cyclodextrin glycosyltransferase (CGTase) with cyclization activity of 43.5 ± 4.4 U/L in M1 medium containing 1% starch and 6% NaCl in nutrient broth at 37 ºC, pH 9.0, after 48h. This is the first report of CGTase from this bacterium. 0.1% starch was found to induce CGTase, and further optimization using one variable at a time approach followed by statistical optimization led to 5.5-fold enhancement resulting in 240.5 ± 5.46 U/L. Six parameters were identified as positive signals using Plackett-Burman (PB). Of these, yeast extract, MgSO4 and tryptone were taken further for Response Surface Methodology (RSM) by disposing beef extract and fixing starch and soya peptone. The optimized M4 medium consisted of tryptone (0.1%, w/v), yeast extract (0.25%, w/v), MgSO4 (8mM, w/v), potato starch (0.1%, w/v) and soya peptone (0.2%, w/v). CGTase was further purified with 6.44-fold purification and 19.32% yield employing starch affinity. It was found to be monomeric, corresponding to a size of 68kDa as estimated by SDS-PAGE and was further confirmed to be 65kDa by size exclusion chromatography. γ-Cyclodextrins were produced as the major product with a conversion of 5% soluble starch into 20.38% γ-cyclodextrins after 24h reaction, as determined by HPLC. Peptide fingerprint after LC-MS analysis matched with IPT/TIG domain-containing protein within the genome of E. caseinilytica. Further blastp analysis revealed the closest homology with γ-CGTase from an alkalophilic E. clarkii, thereby confirming CGTase from E. caseinilytica as γ-CGTase.

Role of extracellular polymeric substances in leaching and bioconcentration of benzophenone-3 from microplastic fragments

Adverse effects of microplastics (MPs) are exacerbated by plastic additives such as benzophenone-3 (BP-3). The aim of the present study was to evaluate the role of extracellular polymeric substances (EPS) of Chlorella vulgaris in leaching BP-3 additive (3.0 ± 0.2% wt/wt) from polyethylene MP fragments (99.8 ± 4.1 µm) and subsequent bioconcentration in Daphnia magna. BP-3 leaching in M4 medium was higher at pH 8 than at pH 6, because of the higher solubility of BP-3 (pKa=7.07) at pH 8. However, EPS reduced BP-3 leaching in M4 medium, possibly because of repulsive interactions between the negatively charged EPS and anionic BP-3. Thus, BP-3 leaching was greater at lower pH (6 >8) and EPS concentration (20 >50 mg L−1 as total organic carbon), which was well related to BP-3 sorption capacity of EPS. Although BP-3 uptake in D. magna was decreased at pH 8 by increasing EPS concentration, the bioconcentration of BP-3 in D. magna was increased, possibly because of reduced BP-3 elimination. These findings suggest the important role of EPS in the bioconcentration of anionic plastic additives, which should be further evaluated to understand the underlying toxicokinetic mechanisms.

Effect of growing media on the germination and seedling growth of Borneo sour eggplant

Borneo sour eggplant, a popular indigenous fruit vegetable in Borneo, has become an important fruit not only to local people, but also to tourists who visit Borneo. One of the challenges in planting this crop is lacking information, particularly in finding suitable media for its growth, eventually inhibiting local farmers from growing the crop commercially. This study was conducted to investigate the effects of 4 growing media compositions (M1: topsoil, M2: topsoil and compost, M3: topsoil and cocopeat and M4: topsoil, compost and cocopeat) on the germination and growth performance of sour eggplant seedlings. The experiment was tested in a randomized complete block design (RCBD) with 10 replicates. At the end of germination period, seeds planted in M4 medium produced the highest germination rate of 96.67% and was significantly different (P &lt; 0.05) from those sown in other growing media. However, the performance of the seedlings in terms of height, stem diameter, number of leaves and length of leaves showed significant increase (P &lt; 0.05) in M2 medium. This study has demonstrated that M2 medium could improve the growth performance of the seedlings and therefore is recommended as suitable growing media for planting sour eggplant. Keywords: Growth performance, growing medium, indigenous fruit, sour eggplant.

Isolation, distribution and evaluation of cytotoxic and antioxidant activity of cultivable actinobacteria from the Oman Sea sediments

Screening bioactive natural products from bacteria is a determinative step in the drug discovery programs. The present study aim to isolate actinobacteria from the Oman Sea sediments for determining the effects of different culture media and treatments on the yield of the isolation process, and measure the DPPH radical scavenging and Artemia cytotoxic activity of culture extracts of the actinobacterial isolates. A total of 290 actinobacterial isolates were collected from 14 sediment samples. Heat treatment (40.68%) and M4 medium (29.31%) exhibited the maximum isolation rates of actinobacteria. Streptomyces isolates were dominantly distributed in all of the investigated stations according to 16S rRNA gene sequencing. The distribution pattern of Streptomyces followed a depth-dependent frequency trend, whereas the members of rare genera including Micromonospora, Nocardia Actinoplanes, Nocardiopsis, Saccharopolyspora and Crossiella were distributed in deeper stations. Approximately, 25% of the examined isolates could scavenge 90% of 10–4 mol/L DPPH solutions at 1 250 µg/mL final concentration of their ethyl acetate culture extracts. Furthermore, the most potent extracts could scavenge DPPH radicals with IC50 ranges from 356.8 to 566.4 µg/mL. Brine shrimp cytotoxicity tests showed that 38.88% of the examined culture extracts exhibited LC50 lower than 1 000 µg/mL against the Artemia cells. Moreover, the most potent culture extracts exhibited LC50 range from 335.4 to 534.4 µg/mL. Phylogenetic analysis by 16S rRNA gene sequence revealed that the OS 005, OS 263 and OS 157 closely related to Streptomyces djakartensis, Streptomyces olivaceus and Nocardiopsis dassonvillei respectively. These results suggested the widespread distribution of the antioxidant and cytotoxic producing actinobacteria in the Oman Sea sediments, which could be considered as promising candidates for the discovery of microbial bioactive compounds.

Changes in the Profiling of Fatty Acids of Glycine max L. (Soybean) Callus after Mutagen Treatments

THE AIM of present study was to produce healthy friable calli from soybean cultivar Giza 111 and studying the effect of sodium azide (SA) and ethidium bromide (EtBr) mutagens on its fatty acids (FAs) profile. Seeds were surface sterilized with clorox and ethanol, then cultured on solid Murashige and Skoog medium (MS) to produce 14-days-old in vitro plantlets. Callus induction from hypocotyl (H), cotyledon (C), epicotyl (E) and leaf (L) explants were tested using different combinations of 2,4-dichlorophenoxy acetic acids (2,4-D) and N6-benzyladenine (BA). M4 (MS+ 0.5mg/L BA) medium was found to be the supreme medium for callus proliferation. Both H-and E-explants were the best explants for this proliferation. Growth dynamics of H- and E-calli was established. The best harvesting time for both calli was the 4th week. H-calli were the most rapidly growing one. The concentrations 0.5, 1.0 and 1.5mM of each mutagen with two soaking times (½ and 1hr) were used for the treatment of H-calli. After mutagenic treatment, treated-H-calli were cultured on callus induction media. A month later, FAs profiling of mutated and control H-calli were performed using gas liquid chromatography (GLC). The Fw pattern of SA and EtBr-treated-H-calli were similar to the control one but with lower values. Saturated (SFAs) and unsaturated-fatty acids (UFAs) contents were raised up by increasing the concentrations of SA and time of incubation. In EtBr-treated-H-calli, marked elevations of SFAs over UFAs were recorded exhibiting a switch in their balance by increasing concentrations and soaking times.

729. Improved Detection of Adenovirus with the FilmArray Respiratory Panel 2 Panel in a Pediatric Population

BackgroundHuman adenovirus (ADV) respiratory infections are associated with up to 8% of all identified viral causes of acute respiratory illnesses, especially among young children. They are therefore included as part of multiplex syndromic respiratory testing. Earlier versions of the FilmArray Respiratory Panel were targeted to genogroups B, C, and E. However, all adenovirus genogroups are associated with disease particularly in immunocompromised patients. Recently, a new version of the FilmArray Respiratory Panel 2 (RP2) has been FDA cleared with significant modifications to the adenovirus assay. The goal of this study was to compare the RP2 adenovirus assay to that of the previous version (RP1.7) and our laboratory-developed (LD) adenovirus PCR.MethodsAnalytical comparison: Ten stocks of know adenovirus serotypes representing genogroups A-F were diluted in M4 media and tested on RP2, RP1.7 and LD PCR to determine the relative limits of detection (LOD). Clinical comparison: A total of 423 pediatric nasopharyngeal samples were tested using RP2, RP1.7, adenovirus LD PCR. In addition we performed genotyping PCRs on most adenovirus positive samples based on the availability.ResultsAnalytical evaluation revealed that for the 10 serotypes (18, 32, 7, 14, 5, 6, 20, 29, 4 and 40), RP2 showed at least 100-fold increase in sensitivity for six serotypes representing genogroups A, D and F. For B, C and E, the relative LODs were comparable.In the 423 clinical samples, there was an overall agreement of 94% between RP2 and RP1.7. Among those RP2+/RP1.7− samples (n = 20), 17 samples were confirmed ADV positive by LD PCR. They all had low viral burden (Ct values >30), among them nine samples had sole detections of adenovirus types A, D and F. Five samples were RP2-/RP1.7+, two samples were confirmed by LD PCR and both were type C. Overall, there was a 22% increase in Adenovirus with RP2 compared with RP1.7.ConclusionThe RP2 adenovirus assay has enhanced inclusivity and lower LOD for all adenovirus genogroups in comparison to RP1.7. Coupled with its faster run time and additional targets, RP2 represents a significant improvement for the syndromic detection of respiratory infections.Disclosures A. Leber, Nationwide Children’s Hospital: Research Contractor, Research support.

Effects of straw leachates from Cry1C-expressing transgenic rice on the development and reproduction of Daphnia magna

The transgenic rice line T1C-19 provides high resistance to lepidopteran pests because of the synthesis of the Bacillus thuringiensis (Bt) insecticidal protein Cry1C. It thus shows good prospect for commercial planting in China. Species of Cladocera, an order of aquatic arthropods commonly found in aquatic ecosystems such as rice paddies, might be exposed to the insecticidal protein released from Bt-transgenic rice-straw residues. For the study reported herein, we used Daphnia magna (water flea) as a representative of Cladocera to evaluate whether aquatic arthropods are adversely affected when exposed to Bt rice-straw leachates. We exposed D. magna to M4 medium containing various volume percentages of medium that had been incubated with T1C-19 rice straw or rice straw from its non-transformed near-isoline Minghui 63 (MH63) for 21 days. Compared with pure M4 medium (control), the fitness and developmental and reproduction parameters of D. magna decreased significantly when exposed to rice-straw leachates; conversely, no significant differences between the T1C-19 and MH63 rice-straw leachate treatments were observed, indicating that the Bt rice straw leachate did not adversely affect this non-target species.