Rotavirus surveillance in Pakistan during 2015-2016 reveals high prevalence of G12P[6].

The G12 rotavirus genotype has emerged globally since their first detection in 1987 from the Philippines; however it remains a rare cause of gastroenteritis in Pakistan. Rotavirus surveillance conducted during 2015-2016, assessed 3446 children <5 years hospitalized for gastroenteritis and found 802 (23.2%) positive on ELISA. Genotyping of a subset of positive samples (n = 319) revealed G12P[6] (11.28%) as the third most common G/P combination following G3P[8] (28.5%) and G1P[8] (12.5%); G2P[4] (10.65%) and G3P[6] (8.15%) were other frequently detected strains. Phylogenetic analysis of G12 strains from Pakistan revealed high genetic similarity to G12 strains from Italy, Thailand, Korea, and Great Britain as well as local strains within G12 lineage III. In conclusion, G12P[6] was a major contributor of RVA gastroenteritis in Pakistani children. Robust surveillance after the introduction of rotavirus vaccines will help determine the evolution of G12 and other circulating genotypes in the country.

Caffeic acid derivatives production by hairy root cultures of Echinacea purpurea.

Inoculation of leaf explants of Echinacea purpurea (Moench) with Agrobacterium rhizogenes induced hairy roots with the capacity to produce biologically active caffeic acid derivatives (CADs), especially cichoric acid. The kinetics of growth, the uptake of macronutrients, and the accumulation of CADs were investigated in heterotrophically cultured hairy roots for a 50 day period. A maximum of 12.2 g L(-1) dry biomass was achieved in MS nutrients supplemented with 30 g L(-1) sucrose on day 40. The mathematical relationship between hairy root growth and conductivity was established during the exponential phase in Erlenmeyer flasks. HPLC analyses of methanolic (0.1% phosphoric acid; 70:30, v/v) extracts from hairy roots revealed the presence of important CADs: cichoric acid (19.21 mg g(-1) dry biomass), caftaric acid (3.56 mg g(-1) dry biomass), and chlorogenic acid (0.93 mg g(-1) dry biomass). These results demonstrate that biotechnological production of CADs in hairy roots of E. purpurea is possible. Furthermore, these hairy root cultures offer, for the very first time, an excellent biological model to study the biosynthetic pathway of medicinally important CADs.

Gibberellic acid increases secondary metabolite production in Echinacea purpurea hairy roots.

Gibberellic acid (GA(3)) is reported to have diverse effects on hairy root cultures of many plant species; therefore, the effects of GA(3) on the growth, secondary metabolite production (caffeic acid derivatives and lignin), phenylalanine ammonia lyase (PAL) activity, and free radical scavenging activity of light-grown Echinacea purpurea L. hairy roots were investigated. Eight concentrations of GA(3), ranging from 0.005 to 1.0 µM, were added to shake flask cultures. The moderate GA(3) concentration, 0.025 µM, resulted in the highest concentrations of cichoric acid, caftaric acid, and chlorogenic acid, as well as increased PAL activity, cell viability, and free radical scavenging activity, while higher and lower GA(3) concentrations resulted in reduced levels compared to the control (lacking GA(3)). The moderate GA(3) concentration also affected root morphogenesis; supplementation with 0.025 µM GA(3) resulted in the development of thick, dense, purple-colored roots, while roots exposed to the higher and lower concentrations of GA(3) were thin and off-white. This study demonstrates that supplementation with GA(3) may be an excellent strategy to optimize the production of secondary metabolites from E. purpurea hairy root cultures; however, the GA(3) concentration is a critical factor.

In vitro larvicidal potential against Anopheles stephensi and antioxidative enzyme activities of Ginkgo biloba, Stevia rebaudiana and Parthenium hysterophorous.

OBJECTIVE: To investigate in vitro larvicidal and antioxidant enzymes potential of the medicinal plants Ginkgo biloba (G. biloba), Stevia rebaudiana (S. rebaudiana) and Parthenium hysterophorous (P. hysterophorous) against Anopheles stephensi (An. stephensi) 4th instars larvae. METHODS: For evaluation of larvicidal potential, the ethanolic, methanolic and dichloromethane leaves extracts of three different plants were used in dose-dependent experiments in two media, while the antioxidant enzymes activities were investigated using four different methods viz., superoxide dismutase, peroxidase, ascorbate and catalase. RESULTS: An. stephensi has developed resistance to various synthetic insecticides, making its control increasingly difficult. The comparative performance of ethanolic extracts (65%-90%) was found better than the methanolic extract (70%-87%) and dichloromethane extract (60%-70%). Among the three plants extracts tested in two media, S. rebaudiana exhibited higher larvicidal activity with LC(50) (24 h) in methanolic extract than P. hysterophorous and G. biloba. G. biloba and P. hysterophorous exhibited the strongest antioxidative enzymes activity and S. rebaudiana were less active and no significant difference was observed. CONCLUSIONS: These three plants exhibit larvicidal potential and can be further used for vector control alternative to synthetic insecticide due to eco-friendly and diseases control, furthermore these plant species have potent antioxidative enzyme activities, therefore, making them strong natural candidate particularly for diseases which are caused due to free radicals.

Light-enhanced caffeic acid derivatives biosynthesis in hairy root cultures of Echinacea purpurea.

Light plays an important role in almost all plant developmental processes and provides the fundamental building blocks for growth, development, primary and secondary metabolism. The effects of light on growth rate and caffeic acid derivative (CADs) biosynthesis in hairy root cultures of Echinacea purpurea (Moench) were assessed. Light-grown hairy roots accumulated increased levels of anthocyanins, which became visible in outer cell layer of the cortex as a ring of purple color. The light-grown root cultures also had radially thickened morphology compared with the dark-grown controls. The growth rate and cell viability of the hairy root cultures in light did not show obvious difference in comparison with those in dark. However, biosynthesis of CADs including cichoric acid, caftaric acid, chlorogenic acid and caffeic acid was significantly increased in hairy root cultures grown in the light. The enhanced accumulation of CADs and anthocyanins in E. purpurea hairy root cultures was correlated to an observed light-stimulated activity of phenylalanine ammonium lyase (PAL).