Molecular characterization and marker based chemotaxonomic studies of Podophyllum hexandrum Royle.

Detailed chemical studies and RAPD analysis were done in different populations of Podophyllum hexandrum collected from high altitude regions of North Western Himalayas. Random amplified polymorphic DNA (RAPD) analysis revealed a high degree of genetic diversity among the 12 collected accessions, attributed to their geographical and climatic conditions. HPLC analysis also revealed variation in the concentration of two major marker compounds which lead to the identification of a chemotype. The study demonstrated that RAPD and chemical markers are very useful tools to compare the genetic relationship and pattern of variation among such prioritized and endangered medicinal plants.

Comparative studies and identification of camptothecin produced by an endophyte at shake flask and bioreactor.

The fungus showing homology with Nodulisporium by 28S ribosomal gene sequencing, which has been discovered as an endophyte on medicinal plant Nothapodytes foetida, was found to produce 45 and 5.5 microg of camptothecin (CPT) per gram of mycelia at bioreactor and at shake flask, respectively, which was further quantified and characterised by various spectroscopic analyses.

Anthelmintic activity of extracts of Artemisia absinthium against ovine nematodes.

The increasing prevalence of anthelmintic resistant strains of helminths, drug residues in animal products and high cost of conventional anthelmintics has created an interest in studying medicinal plants as an alternative source of anthelmintics. Artemisia absinthium Linn. (Tethwen) is used traditionally by people as a vermifuge in addition to its other livestock uses. The objective of this study was to evaluate the anthelmintic efficacy of crude aqueous extracts (CAE) and crude ethanolic extracts (CEE) of the aerial parts of A. absinthium in comparison to albendazole against the gastrointestinal (GI) nematodes of sheep. To fulfill the objectives, the worm motility inhibition assay was utilized in order to investigate the direct effects of plant extracts on the survival of the adult nematodes under in vitro conditions and faecal egg count reduction assay to investigate the effects on faecal egg output of GI nematodes under in vivo conditions. Significant anthelmintic effects of CAE and CEE on live adult Haemonchus contortus worms (P < 0.005) were observed in terms of the paralysis and/or death of the worms at different hours post-treatment (PT), however, CEE were more efficacious than CAE. The oral administration of the extracts in sheep was associated with significant reduction in faecal egg output by the GI nematodes. The CEE was as effective as the reference drug-albendazole and demonstrated faecal egg count reduction (FECR) of 90.46% in sheep at 2.0 g kg(-1) body weight on day 15 PT followed by 82.85% FECR at 1.0 g kg(-1) bw on day 15 PT. The CAE showed less activity and resulted in maximum of 80.49% FECR at 2.0 g kg(-1)bw. Dosage had a significant (P < 0.05) influence on the anthelmintic efficacy of A. absinthium. The better activity of CEE can be attributed to the greater concentration of alcohol soluble active anthelmintic principle/s and a more rapid transcuticular absorption of the CEE into the body of the worms when compared with the CAE. The results of the present study suggest that A. absinthium extracts are a promising alternative to the commercially available anthelmintics for the treatment of GI nematodes of sheep.

Epidemiology of gastrointestinal nematodes of sheep managed under traditional husbandry system in Kashmir valley.

The present study was conducted with the objective to investigate the seasonal epidemiological prevalence of gastrointestinal tract (GIT) nematodes in different age groups, sexes and breeds (genotypes) of sheep through necropsy and faecal analysis over a period of 2 years in Kashmir valley, India. A total of 1533 sheep were examined [faecal examination: 1035 (year 1: 561, year 2: 474); necropsy: 498 (year 1: 232, year 2: 266)]. Out of these, 945 (61.64%) were found infected [faecal examination: 697 (67.34%, year 1: 390 (69.51%), year 2: 307 (46.99%); necropsy: 248 (49.79%, year 1: 123 (53.01%), year 2: 125 (64.69%)] with GIT nematodes. The over all prevalence of GIT nematodes in sheep in year 1 was 64.76 and 58.37% in year 2 (P=0.04). The parasites in decreasing order of prevalence (%) in sheep were Haemonchus contortus (59.6); Ostertagia circumcincta (38.0); Bunostomum trigonocephalum (37.7); Chabertia ovina (37.7); Trichostrongylus spp. (33.9); Nematodirus spathiger (29.4); Oesophagostomum columbianum (28.4); Trichuris ovis (23.5) and Marshallagia marshalli (22.1). Season, sex, age, and genotype were the factors that influenced the epidemiological prevalence of GIT nematodes in sheep in the present study. The maximum nematode infection was observed in summer season and lowest in winter (P=0.0005). Local Kashmiri breed was less infected as compared to other genotypes (P>0.05). Lower age groups were more infected than adult animals (P>/=0.05). Prevalence was higher in rams (males) than eves (females) (P>0.05). The present study will initially be of great significance to add to the existing knowledge of the epidemiology of GIT nematodes of small ruminants and the findings will be quite helpful to devise the appropriate control and prophylactic strategies for GIT nematodiasis of sheep reared under the temperate agro-climatic conditions.

An endophytic Neurospora sp. from Nothapodytes foetida producing camptothecin.

The medicinal plant, Nothapodytes foetida contains a number of important alkaloids like camptothecin (an anticancer drug molecule) but its concentration is less to meet the existing demand of this important molecule, so in an effort for accessible availability of camptothecin. An endophyte (designated ZP5SE) was isolated from the seed of Nothapodytes foetida and was examined as potential source of anticancer drug lead compound i.e. camptothecin, when grown in Sabouraud liquid culture media under shake flask conditions. The presence of anticancer compound (camptothecin) in this fungus was confirmed by chromatographic and spectroscopic methods in comparison with authentic camptothecin. Isolated endophyte (Neurospora crassa) producing camptothecin may become an easily accessible source for the production of precursor anticancer drug molecule in future at large scale.

Anthelmintic efficacy of Achillea millifolium against gastrointestinal nematodes of sheep: in vitro and in vivo studies.

Achillea millifolium Linn., commonly called 'Pehl-ghasa', is used by farmers in traditional animal health care as a vermifuge. The objective of this study was to evaluate the anthelmintic efficacy of crude aqueous extracts and crude ethanolic extracts of entire A. millifolium against the gastrointestinal nematodes of sheep. The worm motility inhibition assay was used for in vitro studies and faecal egg count reduction assay was used for in vivo studies. In vitro studies revealed significant anthelmintic effects of aqueous extracts and ethanolic extracts on live Haemonchus contortus worms (P < 0.05) as evident from their paralysis and/or death at 8 h post exposure. Aqueous extracts of A. millifolium resulted in a mean worm motility inhibition of 94.44%, while ethanolic extracts resulted in mean worm motility inhibition of 88.88%. The mean mortality index of aqueous extracts was 0.95 while for ethanolic extracts it was 0.9. The lethal concentration 50 was 0.05 mg ml(-1) for aqueous extracts and 0.11 mg ml(-1) for ethanolic extracts. The in vivo anthelmintic activity of aqueous and ethanolic extracts of A. millifolium demonstrated a maximum (88.40%) nematode egg count reduction in sheep treated with aqueous extracts at 2 g kg(-1) body weight on day 15 after treatment. Ethanolic extracts resulted in a maximum of 76.53% reduction in faecal egg counts on day 15 after treatment with 2 g kg(-1) body weight. Thus, the aqueous extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions, and could be due to the presence of water-soluble active principle/s in A. millifolium. It is concluded that the entire plant of A. millifolium possesses significant anthelmintic activity and could be a potential alternative for treating cases of helminth infections in ruminants.

The epidemiology of paramphistomosis of sheep (Ovis aries L.) in the north west temperate Himalayan region of India.

An epidemiological study with the objective to assess the prevalence of paramphistomosis in association with season, age, sex and breed was carried out in naturally infected sheep over a period of two years from February 2005 to January 2007. Gastrointestinal tract (GIT) and faecal examination were conducted monthly to monitor the seasonal occurrence of paramphistomosis. 793 sheep were examined in the first year, out of which 7.06% were positive for Paramphistomum infection. In the second year, 740 animals were investigated and 7.7% were infected. The overall prevalence of paramphistomosis was 7.3% with a mean of 56.50 +/- 0.50 and 95% confidence interval (CI) (lower bound: 50.1469; upper bound: 62.8531). The prevalence of paramphistomosis through GIT examination (P = 0.593) was 7.6% at 95% CI (lower bound: -19.1186; upper bound: 57.1186) and the prevalence through faecal examination (P = 0.884) was 7.2% at 95% CI (lower bound: 5.7345; upper bound: 69.2655). Generally, season and age were the factors found to have a significant influence on the risk of paramphistomosis in sheep. The highest infection was found in the summer season (P < 0.005); lower age groups (P < 0.005) in males and in migratory (Bhakarwal) breed (P >or= 0.005). Winter, adult animals, females and local breed reported low infection. The present study will be of great significance to understand the epidemiology of gastrointestinal helminthes of sheep initially in the resource poor communities of Himalayan region and will definitely be helpful to devise appropriate control strategies for paramphistomosis.

Evaluation of anthelmintic activity of Iris hookeriana against gastrointestinal nematodes of sheep.

The objective of this study was to evaluate the anthelmintic efficacy of Iris hookeriana Linn. rhizome against gastrointestinal nematodes of sheep. A worm motility inhibition assay was used for in vitro study and a faecal egg count reduction assay was used for an in vivo study. The in vitro study revealed anthelmintic effects of crude aqueous extracts and crude ethanolic extracts on live Trichuris ovis worms (P < or = 0.05) as evident from their paralysis and/or death at 8 h after exposure. The aqueous extracts of I. hookeriana resulted in a mean worm motility inhibition of 54.0%, while ethanolic extracts resulted in a mean worm motility inhibition of 84.6%. The mean mortality index of aqueous extracts was 0.55, while for ethanolic extracts it was 0.85. The lethal concentration 50 for aqueous extracts was 0.45 mg ml- 1 and for ethanolic extracts it was 0.15 mg ml- 1. The in vivo anthelmintic activity of aqueous and ethanolic extracts of I. hookeriana in sheep naturally infected with mixed species of gastrointestinal nematodes demonstrated a maximum (45.62%) egg count reduction in sheep treated with ethanolic extracts at 2 g kg- 1 body weight on day 10 after treatment, closely followed by ethanolic extracts at 1 g kg- 1 body weight on day 10 after treatment (43.54% egg count reduction). The aqueous extracts resulted in a maximum of 31.53% reduction in faecal egg counts on day 10 after treatment with 1 g kg- 1 body weight. Thus ethanolic extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions; this could be due to the presence of alcohol-soluble active ingredients in I. hookeriana. From the present study it can be suggested that I. hookeriana rhizome exhibited significant anthelmintic activity against gastrointestinal nematodes of sheep and has the potential to contribute to the control of gastrointestinal nematode parasites of small ruminants.

Podophyllum hexandrum Offers Radioprotection by Modulating Free Radical Flux: Role of Aryl-Tetralin Lignans.

We have evaluated the effect of variation in aryl-tetralin lignans on the radioprotective properties of Podophyllum hexandrum. Two fractionated fractions of P. hexandrum [methanolic (S1) and chloroform fractions (S2)], with varying aryl-tetralin lignan content were utilized for the present study. The peroxyl ion scavenging potentials of S1 and S2 were found to be comparable [i.e. 45.88% (S1) and 41% (S2)] after a 48 h interval in a time-dependent study, whereas in a 2 h study, S2 exhibited significant (P < 0.05) antioxidant activity in different metal ion + flux states. In the aqueous phase, S2 exhibited non-site-specific reactive oxygen species scavenging activity, i.e. 73.12% inhibition at 500 mug ml(-1). S1 exhibited 58.40 +/- 0.8% inhibition (at 0.025 mug ml(-1)) of the formation of reactive nitrite radicals, comparable to S2 (52.45 +/- 0.825%), and also showed 45.01% site-specific activity (1000 mug ml(-1)), along with significant (P < 0.05) electron donation potential (50-2000 mug ml(-1)) compared to S2. Such activities of S1 could be attributed to the significantly (P < 0.05) higher levels of podophyllotoxin beta-d-glucopyranoside (16.5 times) and demethyl podophyllotoxin glucoside (2.9 times) compared with S2. Together, these findings clearly prove that aryl-tetralin lignan content influences the radiation protective potential of the Podophyllum fractions to a great extent.

Preparative high-performance liquid chromatographic separation of naphthodianthrones from St. John's Wort.

St. John's Wort (Hypericum perforatum), a perennial flowering plant, has been used medicinally for thousands of years and has most recently been identified as an effective treatment for mild to moderate depression and neuralgic disorders. This work presents a procedure for the isolation of naphthodianthrones from St. John's Wort by an accelerated extraction and separation of marker compounds by preparative high-performance liquid chromatography (HPLC) with photodiode array detection. The accelerated extraction method minimizes the extraction time and increases the yield, and the marker compounds obtained by preparative HPLC are of 98% purity. The compounds are characterized by liquid chromatography-mass spectrometry (electrospray ionization) and NMR spectra.

A brevifoliol analogue from the Himalyan yew Taxus wallichiana.

The needles of Taxus wallichiana gave a new brevifoliol derivative whose structure was established by spectral data.

Volatile Constituents of Dracocephalum nutans.

The essential oil of DRACOCEPHALUM NUTANS L. (Lamiaceae) has been investigated for the first time. The GC-MS analysis of the oil showed the presence of about 25 monoterpenoids including pinocamphone (56.4%), beta-pinene (12.7%), isopinocamphone (4.3%), alpha-phellandrene (4.6%), and isopinocampheol (3.7%).