Angelica archengelica extract induced perturbation of rat skin and tight junctional protein (ZO-1) of HaCaT cells

Background and purpose of the study: Herbal enhancers compared to the synthetic ones have shown less toxis effects. Coumarins have been shown at concentrations inhibiting phospoliphase C-Y (Phc-Y) are able to enhance tight junction (TJ) permeability due to hyperpoalation of Zonolous Occludense-1 (ZO-1) proteins. The purpose of this study was to evaluate the influence of ethanolic extract of Angelica archengelica (AA-E) which contain coumarin on permeation of repaglinide across rat epidermis and on the tight junction plaque protein ZO-1 in HaCaT cells. Methods: Transepidermal water loss (TEWL) from the rat skin treated with different concentrations of AA-E was assessed by Tewameter. Scanning and Transmission Electron Microscopy (TEM) on were performed on AA-E treated rat skin portions. The possibility of AA-E influence on the architecture of tight junctions by adverse effect on the cytoplasmic ZO-1 in HaCaT cells was investigated. Finally, the systemic delivery of repaglinide from the optimized transdermal formulation was investigated in rats. Results: The permeation of repaglinide across excised rat epidermis was 7-fold higher in the presence of AA-E (5% w/v) as compared to propylene glycol:ethanol (7:3) mixture. The extract was found to perturb the lipid microconstituents in both excised and viable rat skin, although, the effect was less intense in the later. The enhanced permeation of repaglinide across rat epidermis excised after treatment with AA-E (5% w/v) for different periods was in concordance with the high TEWL values of similarly treated viable rat skin. Further, the observed increase in intercellular space, disordering of lipid structure and corneocyte detachment indicated considerable effect on the ultrastructure of rat epidermis. Treatment of HaCaT cell line with AA-E (0.16% w/v) for 6 hrs influenced ZO-1 as evidenced by reduced immunofluorescence of anti-TJP1 (ZO-1) antibody in Confocal Laser Scanning Microscopy studies (CLSM) studies. The plasma concentration of repaglinide from transdermal formulation was maintained higher and for longer time as compared to oral administration of repaglinide. Major conclusion: Results suggest the overwhelming influence of Angelica archengelica in enhancing the percutaneous permeation of repaglinide to be mediated through perturbation of skin lipids and tight junction protein (ZO-1).

Ethnobotany of the Monpa ethnic group at Arunachal Pradesh, India

Background: The present paper documents the uses of plants in traditional herbal medicine for human and veterinary ailments, and those used for dietary supplements, religious purpose, local beverage, and plants used to poison fish and wild animals. Traditional botanical medicine is the primary mode of healthcare for most of the rural population in Arunachal Pradesh. Materials and methods: Field research was conducted between April 2006 and March 2009 with randomly selected 124 key informants using semi-structured questionnaire. The data obtained was analyzed through informant consensus factor (F(IC)) to determine the homogeneity of informant's knowledge on medicinal plants. Results: We documented 50 plants species belonging to 29 families used for treating 22 human and 4 veterinary ailments. Of the medicinal plants reported, the most common growth form was herbs (40%) followed by shrubs, trees, and climbers. Leaves were most frequently used plant parts. The consensus analysis revealed that the dermatological ailments have the highest F(IC) (0.56) and the gastro-intestinal diseases have F(IC) (0.43). F(IC) values indicated that there was high agreement in the use of plants in dermatological and gastro-intestinal ailments category among the users. Gymnocladus assamicus is a critically rare and endangered species used as disinfectant for cleaning wounds and parasites like leeches and lice on livestocks. Two plant species (Illicium griffithii and Rubia cordifolia) are commonly used for traditional dyeing of clothes and food items. Some of the edible plants recorded in this study were known for their treatment against high blood pressure (Clerodendron colebrookianum), diabetes mellitus (Momordica charantia), and intestinal parasitic worms like round and tape worms (Lindera neesiana, Solanum etiopicum, and Solanum indicum). The Monpas of Arunachal Pradesh have traditionally been using Daphne papyracea for preparing hand-made paper for painting and writing religious scripts in Buddhist monasteries. Three plant species (Derris scandens, Aesculus assamica, and Polygonum hydropiper) were frequently used to poison fish during the month of June-July every year and the underground tuber of Aconitum ferrox is widely used in arrow poisoning to kill ferocious animals like bear, wild pigs, gaur and deer. The most frequently cited plant species; Buddleja asiatica and Hedyotis scandens were used as common growth supplements during the preparation of fermentation starter cultures. Conclusion: The traditional pharmacopoeia of the Monpa ethnic group incorporates a myriad of diverse botanical flora. Traditional knowledge of the remedies is passed down through oral traditions without any written document. This traditional knowledge is however, currently threatened mainly due to acculturation and deforestation due to continuing traditional shifting cultivation. This study reveals that the rural populations in Arunachal Pradesh have a rich knowledge of forest-based natural resources and consumption of wild edible plants is still an integral part of their socio-cultural life. Findings of this documentation study can be used as an ethnopharmacological basis for selecting plants for future phytochemical and pharmaceutical studies.

Cytotoxic and pro-apoptotic effects of Abrus precatorius L. on human metastatic breast cancer cell line, MDA-MB-231

Abrus precatorius is highly regarded as a universal panacea in the herbal medicine with diverse pharmacological activity spectra. This experimental study on the mechanism of the anticancer activity of A. precatorius leaf extracts, may offer new evidence for A. precatorius in the treatment of breast cancer in clinical practice. Cell death was determined by using MTT assay. Further analyses were carried out by doing DNA laddering, PARP cleavage, FACS, semi-quantitative RT-PCR and detection of cellular reactive oxygen species (ROS) by DCFDA assay. A. precatorius showed very striking inhibition on MDA-MB-231 cells. MTT assay showed more than 75 % inhibition of the cells and treated cells indicated visible laddering pattern with thick compact band. PARP cleavage produced 89 kDa cleavage product which was associated with apoptosis. Flow cytometer exhibited a sub-G0/G1 peak as an indicative of apoptosis. mRNA expression level of apoptosis-related genes p21 and p53 was markedly increased in cells treated with the extract as compared to control. The up-regulation of p21 and p53 may be the molecular mechanisms by which A. precatorius extract which induces apoptosis. An increase in the concentration of A. precatorius extract does not generate ROS, instead it reduces ROS formation in MDA-MB-231 cells, as evident from the shift in fluorescence below untreated control. This is the first report showing that A. precatorius leaf extract exhibits a growth inhibitory effect by induction of apoptosis in MDA-MB-231 cells. Our results contribute towards validation of the A. precatorius extract as a potentially effective chemopreventive or therapeutic agent against breast cancer.

Evaluation of the Genotoxic and Antigenotoxic Potential of the Alkaloid Punarnavine from Boerhaavia diffusa

Boerhaavia diffusa is a traditional herbal medicine extensively used in the Ayurveda and Unani forms of medicine in India and many parts of the world. Different parts of the plant are used as an appetizer, alexiteric, eye tonic, for flushing out the renal system, and to treat blood pressure. This study was conducted to evaluate the in vivo genotoxic and/or antigenotoxic potential of punarnavine, a separated alkaloid from the root of B. diffusa using toxicity studies (OECD guideline 474, 1997). The genotoxic and antigenotoxic potential of punarnavine was assayed using the comet assay on lymphocytes, liver, spleen, brain, and bone marrow as well as using the micronucleus test in bone marrow cells including the in vitro chromosomal aberration test. The results demonstrated that none of the tested doses of punarnavine showed genotoxic effects by the comet assay, or clastogenic effects in the micronucleus test. On the other hand, for all cells evaluated, the three tested doses of punarnavine promoted inhibition of DNA damage induced by cyclophosphamide. Based on these results, we concluded that punarnavine, an alkaloid from the Boerhaavia diffusa root, has no genotoxic or clastogenic effects in our experimental conditions. However, it caused a significant decrease in DNA damage induced by cyclophosphamide. It is suggested that the antigenotoxic properties of this alkaloid may be of great pharmacological importance and beneficial for cancer prevention.

In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells

Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l(-1) IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13) and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW) and Echioidinin (ED). Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time-and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.

A glucose-centric perspective of hyperglycemia

Digestion of food in the intestines converts the compacted storage carbohydrates, starch and glycogen, to glucose. After each meal, a flux of glucose (>200 g) passes through the blood pool (4-6 g) in a short period of 2 h, keeping its concentration ideally in the range of 80-120 mg/100 mL. Tissue-specific glucose transporters (GLUTs) aid in the distribution of glucose to all tissues. The balance glucose after meeting the immediate energy needs is converted into glycogen and stored in liver (up to 100 g) and skeletal muscle (up to 300 g) for later use. High blood glucose gives the signal for increased release of insulin from pancreas. Insulin binds to insulin receptor on the plasma membrane and activates its autophosphorylation. This initiates the post-insulin-receptor signal cascade that accelerates synthesis of glycogen and triglyceride. Parallel control by phos-dephos and redox regulation of proteins exists for some of these steps. A major action of insulin is to inhibit gluconeogensis in the liver decreasing glucose output into blood. Cases with failed control of blood glucose have alarmingly increased since 1960 coinciding with changed life-styles and large scale food processing. Many of these turned out to be resistant to insulin, usually accompanied by dysfunctional glycogen storage. Glucose has an extended stay in blood at 8 mM and above and then indiscriminately adds on to surface protein-amino groups. Fructose in common sugar is 10-fold more active. This random glycation process interferes with the functions of many proteins (e.g., hemoglobin, eye lens proteins) and causes progressive damage to heart, kidneys, eyes and nerves. Some compounds are known to act as insulin mimics. Vanadium-peroxide complexes act at post-receptor level but are toxic. The fungus-derived 2,5-dihydroxybenzoquinone derivative is the first one known to act on the insulin receptor. The safe herbal products in use for centuries for glucose control have multiple active principles and targets. Some are effective in slowing formation of glucose in intestines by inhibiting alpha-glucosidases (e.g., salacia/saptarangi). Knowledge gained from French lilac on active guanidine group helped developing Metformin (1,1-dimethylbiguanide) one of the popular drugs in use. One strategy of keeping sugar content in diets in check is to use artificial sweeteners with no calories, no glucose or fructose and no effect on blood glucose (e.g., steviol, erythrytol). However, the three commonly used non-caloric artificial sweetener's, saccharin, sucralose and aspartame later developed glucose intolerance, the very condition they are expected to evade. Ideal way of keeping blood glucose under 6 mM and HbAlc, the glycation marker of hemoglobin, under 7% in blood is to correct the defects in signals that allow glucose flow into glycogen, still a difficult task with drugs and diets.

Um estudo dos textos clássicos do Ayurveda em perspectiva histórico antropológica.

Nas últimas duas décadas a racionalidade ayurvédica tornou-se popular no ociedente e está se expandindo rapidamente. Esta expansão é consequência do renascimetno do Ayurveda na Índia no século XX. Apesae do crescente interesse neste sistema antido de medicina pouco se tem explorado, no nosso meio, da dua gênese histórica e das pesquisas dos textos clássicos, riquíssimos em informação spbre esta antiga medicina e suas ferramentas de diagnóstico e terapêutica prevalente no subcontinente indiano há milhares de anos. O renascimento do Ayurveda se intensificou após a libertação da Índia da dominação britânica em 1947. Na década de 50 vários esforços foram realizados para promover o ensino e desenvolvimento desta racionalidade médica pelo governo indiano. A Medicina Ayurvédica se expandiu rapidamente pelo subcontinente e posteriormente pelo ocidente, Europa e Estados Unidos. No Brasil o Ayurveda chegou a meados dos anos 80 e se desenvolveu principalmente em Goiânia com o Hospital de Medicina Alternativa. Nesta instituição as plantas medicinais brasileiras receberam um leitura da racionalidade ayurvédica através dos vários médicos indianos que lá estiveram. Esta tese de natureza teórico-conceitual, mas com um enfoque histórico antropológico tem como objeto de estudo a gênese do Ayurveda e a análise crítica comparada dos textos clássico nas suas fontes primárias e secundárias. O período de formação desta racionalidade médica na Índia antiga ainda é objeto de muitas discussões dos autores modernos, isto ocorre por que a transformação de uma medicina mágico-religiosa dos textos védicos em um sistema empírico-racional do clássico Ayurveda não foi totalmente esclarecida pelos historiadores e pesquisadores ayurvedisas. Analiseremos os principais textos clássicos e seus autores de uma forma comparativa e simultaneamente tentaremos propor uma gênese histórica do Ayurveda, na antiga Índia, baseada nas traduções das fontes primárias e na literatura secundária dos autores orientais e ocidentais que estiveram ao nosso alcance durante a pesquisa.

Estudo da oferta e produção de atendimentos das práticas integrativas e complementares no SUS no município do Rio de Janeiro

Esta pesquisa é um estudo de caso com abordagem qualitativa e quantitativa, de natureza exploratória que se propõe a analisar as características de oferta e produção das práticas integrativas e complementares, no período de 2006 a 2013 no município do Rio de Janeiro. Como fontes de dados foram utilizados os bancos de dados nacionais sobre oferta de serviços, de profissionais e de produção: o SCNES- Sistema do Cadastro Nacional de Estabelecimentos de Saúde onde são registrados dados da capacidade física e recursos humanos dos estabelecimentos de saúde e o SIASUS Sistema de Informação Ambulatorial onde são registrados os dados da produção ambulatorial do SUS. Optou-se por esses dois bancos de dados por serem ferramentas institucionais de gerenciamento da capacidade instalada e produção utilizadas pelas esferas federal, estadual e municipal. Buscou identificar profissionais cadastrados no SCNES que são autorizados a oferecer práticas integrativas e complementares no município do Rio de Janeiro, detectar os serviços de práticas integrativas e complementares cadastrados no CNES do município do Rio de Janeiro e analisar no Sistema de Informação Ambulatorial, registros de produção em práticas integrativas e complementares do município do Rio de Janeiro. A lista de práticas integrativas e complementares estabelecidas na portaria no 971 de 2006 são homeopatia, medicina tradicional chinesa (onde se inclui a acupuntura), medicina antroposófica, plantas medicinais e fitoterapia, termalismo/crenoterapia. Este estudo proporciona maior visibilidade quanto às características de implementação e institucionalização de uma recente política pública de saúde e contribui com base na análise dos dados encontrados em ferramentas de gestão, para o aprimoramento de ações de acompanhamento e avaliação, estruturação dos serviços, bem como o desenvolvimento sustentável de políticas locais de oferta das terapias complementares do SUS, em consonância com as diretrizes da Política Nacional de Práticas Integrativas e Complementares.

A concessão de patentes de medicamentos fitoterápicos no Brasil: uma análise do período 1997-2004

Este trabalho aborda o problema do consumo desenfreado de produtos naturais no Brasil, especificamente de medicamentos fitoterápicos, com suas consequências, discutindo à luz do Direito de Propriedade Industrial e o Direito Sanitário. Procura relacionar o papel e a responsabilidade do INPI e a ANVISA, órgãos diretamente envolvidos na legislação mencionada, para a melhoria da qualidade de produtos fitoterápicos registrados e lançados no mercado brasileiro. Mostra a necessidade de uma reavaliação dos critérios de concessão de patentes para a área analisada por parte do INPI, bem como uma maior aproximação da ANVISA junto à este órgão, de modo que harmonizem estratégias para que a população tenha acesso a medicamentos fitoterápicos seguros e eficazes. Explicita as estratégias d indústria nacional do setor em alavancar suas vendas no mercado, através de projetos de lei que não traduzem com a realidade preconizada pela OMS, para o uso da Medicina Tradicional/Complementar, contrariando a legislação brasileira para o possível registro dos produtos patenteados. Conclui-se que interesses econômicos, ligados à acirrada concorrência no mercado de fitoterápicos e suplementos nutricionais, induzem à concessão de patentes para tais produtos, em detrimento das diretrizes existentes na Lei de Propriedade Industrial brasileira, em critérios de avaliação.

O sofrimento de adolescentes internados : a escuta psicanalítica na Clínica do Cuidar

A possibilidade de uma intervenção psicanalítica na clínica do cuidar, bem como a escuta de adolescentes cujo sofrimento causado pela doença crônica exige a hospitalização, são as duas questões centrais desta dissertação. Nossa pesquisa tem início na análise da estrutura da clínica psicanalítica que é sustentada na Enfermaria do Núcleo de Estudos da Saúde do Adolescente/NESA/UERJ. Faz-se um percurso pela teoria lacaniana da fase do espelho, como base da identificação especular, para que, em seguida, se possa verificar de que modo o mandamento do amor ao próximo está presente no ato de cuidar. Com Freud, coloca-se em primeiro plano a questão libidinal relativa ao processo da doença física. Destaca-se também o conceito freudiano de inibição que fundamenta a explicação das modalidades de retração da libido. O debate psicanalítico sobre a influência do inconsciente e as alterações que ele provoca no quadro da doença orgânica traz uma contribuição à clínica médica. A ausência de investimento libidinal afetivo nas figuras parentais, na clínica com adolescentes cujo corpo está adoecido, é o ponto em que este trabalho se conclui, demonstrando que para alguns adolescentes esta é a principal razão de seu óbito

Effects of essential oil of Satureja bachtiarica, Satureja khuzestanica and Mentha longifolia on the growth of Actococcus garvieae in rainbow trout fillet

The present research was done with the aim of studing the effects of some medicinal plant essential oils against Lactococcus garvieae and on some spoilage indexes in rainbow trout fillets in 4°C during 18 days. Essential oils of Mentha longifola (20, 40 and 80 μg g-1), Satureja bachtiarica (0.125, 0.25 and 0.50 μg g-1) and Satrureja khuzestanica (0.06, 0.12, 0.25 and 0.50 μg g-1) were used for this study. According to the results, the highest and the lowest microbial load were observed in control group and the groups contained 80 and 40 μg g-1 of Mentha longifola, respectively (P≤0.05). The spoilage indexes were increased after 18 days showing a statistical difference for protein, fat, TVN, FFA and TBA (P≤0.05). The most and the least changes were observed in the the control group and the fillets containing 80 μg g-1 of Mentha longifola, respectively (P≤0.05). The results indicated that, in comparison with control group, using herbal essential oils leads to reduction of Lactococcus garvieae population and spoilage process of the fillets through decreasing in TVN, FFA and TBA. Meanwhile, Mentha longifola having 80 μg g-1 was found to be more effective

Effects of Zataria multiflora Boiss essential oil and nisin on the growth of Staphylococcus aureus in light salted silver carp fillet (Hypophthalmichthys molitrix)

There is an increasing demand in developing newer and safer methods in preserving food products.Among which herbal additives seem to attract evermore attention recently.the major advantage of herbal additives is due to their favorable aroma besides their antimicrobial effects and less expensive than chemical additives. Zataria multiflora Boiss is a native Iranian herb which is used vastly as a food preserver essential oils and also medical usage. Metabolites of harmless bacteria, such as Nisin are also known to be safe preservatives that have antimicrobial activity. However to establish the usefulness of natural antimicrobial preservatives, they must be evaluated alone and in combination with other preservation factors to determine whether there are synergistic effects in rigid media . In this study were evaluated the effects of different concentrations of Zataria multiflora (EO 0, 0.005, 0.015, 0.045, 0.135, 0.405 ,0.810 %) and Nisin(0, 0.15, 0.25, 0.75 μg/ml) and Storage time (up to 21 days) on growth of Staphylococcus aureus ATCC 6538 in a food model system(light salted fish of silver carp, Hypophthalmichthys molitrix). The results on growth of S. aureus were evaluated using SPSS 15.0 statistical software (SPSS 15.0 for windows, SPSS Inc.) and analyzed the logarithm of total count of the bacteria by Tukey Test. Results were considered statistically significant when P≤0.05. The growth of Staphylococcus aureus was affected significantly(P<0.05) by EO and Nisin and also combinations of EO and Nisin. Samples treated with 0.135, 0.405 and 0.810% of thyme essential oil showed a significant decrease on the growth of the bacteria compared with an treated samples(P<0.05). No significant difference was seen on the growth of S.aureus in samples treated with lower concentrations of Z.multiflora(below 0.045%) and untreated group(P>0.05). The most inhibitory effects were seen in samples treated with 0.405% and 0.810% of thyme essential oil until 9 and 12 days after storage,respectively. Also there was significant inhibtory effect(P<0.05) in different concentration of nisin on the organism compared with an treated samples. The synergism effects of the Eo and nisin on the growth rate of the bacteria was significant (P<0.05) compared with untreated samples and samples treated with the Eo or nisin, only. Synergismic effects was observed at concentration of 0.405 and 0.810% of Z. multiflora essential oil with 0.25 μg/ml Nisin, respectively until 15 days after storage. As expected it is preferred to apply the least possible amounts of additives in food preserving that not only are effective and safe but are economically justifiable.

Colony development and physiological characterization of the edible blue-green alga, Nostoc sphaeroides (Nostocaceae, Cyanophyta)

The edible blue-green alga, Nostoc sphaeroides Kutzing, is able to form microcolonies and spherical macrocolonies. It has been used as a potent herbal medicine and dietary supplement for centuries because of its nutraceutical and pharmacological benefits. However, limited information is available on the development of the spherical macrocolonies and the environmental factors that affect their structure. This report described the morphogenesis of N. sphaeroides from single trichomes to macrocolonies. During the process, most structural features of macrocolonies of various sizes were dense maculas, rings, the compact core and the formation of liquid core; and the. laments within the macrocolonies showed different lengths and arrays depending on the sizes of macrocolonies. Meanwhile temperature and light intensity also strongly affected the internal structure of macrocolonies. As microcolonies further increased in size to form 30 mm macrocolonies, the colonies differentiated into distinct outer, middle and inner layers. The. laments of the outer layer showed higher maximum photosynthetic rates, higher light saturation point, and higher photosynthetic effciency than those of the inner layer; whereas the. laments of the inner layer had a higher content of chlorophyll a and phycobiliproteins than those of the outer layer. The results obtained in this study were important for the mass cultivation of N. sphaeroides as a nutraceutical product. (c) 2008 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

Hydrodynamics in an expanded bed of large size ion-exchange resin, and natural product adsorption

Successful applications of expanded bed adsorption (EBA) technology have been widely reported in the literature for protein purification. Little has been reported on the recovery of natural products and active components of Chinese herbal preparations using EBA technology. In this study, the hydrodynamic behavior in an expanded bed of cation resin, 001 x 7 Styrene-DVB, was investigated. Ephedrine hydrochloride (EH) was used as a model natural product to test the dynamic binding capacity (DBC) in the expanded bed. EBA of EH directly from a feedstock containing powdered herbs has also been investigated. These particles are different from commercially available expanded bed adsorbents by virtue of their large size (20S to 1030 gm). When the adsorbent bed is expanded to approximately 1.3 to 1.5 times its settled bed height, the axial liquid-phase dispersion coefficient was found to be of the order 10(-5) m(2) s(-1), which falls into the range 1.0 x 10(-6) to 1.0 X 10(-5) m(2) s(-1) observed previously in protein purification. Because of the favorable column efficiency (low axial dispersion coefficient), the recovery yield and purification factor values of EH directly from a feedstock reached 86.5% and 18, respectively. The results suggest that EBA technology holds promise for the recovery of natural products and active components of Chinese herbal preparations.

Influence of selected Indian immunostimulant herbs against white spot syndrome virus (WSSV) infection in black tiger shrimp, Penaeus monodon with reference to haematological, biochemical and immunological changes

Immunostimulants are the substances, which enhance the non-specific defence mechanism and provide resistance against the invading pathogenic micro-organism. In order to increase the immunity of shrimps against the WSSV, the methanolic extracts of five different herbal medicinal plants like Cyanodon dactylon, Aegle marmelos, Tinospora cordifolia, Picrorhiza kurooa and Eclipta alba were selected and mixed thoroughly in equal proportion. The mixed extract was supplemented with various concentrations viz. 100 (A), 200 (B), 400 (C), and 800 (D) mg kg(-1) through artificial diets individually. The prepared diets (A-D) were fed individually to WSSV free healthy shrimp Penaeus monodon with an average weight of 8.0 +/- 0.5 g for 25 days. Control diet (E), devoid of herbal extract was also fed to shrimps simultaneously. After 25 days of feeding experiment, the shrimps were challenged with WSSV, which were isolated and propagated from the infected crustaceans. The shrimps succumbed to death within 7 days when fed on no herbal immunostimulant diet (E). Among the different concentrations of herbal immunostimulant supplemented diets, the shrimps fed on diet D (800 mg kg(-1)) significantly (P < 0.0001) had more survival (74%) and reduction in the viral load. Also the better performance of haematological, biochemical and immunological parameters was found in the immunostimulant incorporated diets fed shrimps. The present work revealed that the application of herbal immunostimulants will be effective against shrimp viral pathogenesis and they can be recommended for shrimp culture. (c) 2006 Published by Elsevier Ltd.