Estudios de Seguridad Preclínica del extracto hidroalcohólico de semillas de Opuntia soehrensii, una especie alto-andina con actividad anti-Herpes Virus, en ratas Sprague Dawley / Preclinical safety of the hydroalcoholic extract of Opuntia soehrensii seeds with anti-herpes virus activity, in SD rats

Introducción. Alrededor de 3700 millones menores de 50 años con infección por VHS-1 y 491 millones de personas de 15 a 49 años cursan con infección por VHS-2 en el mundo; sus síntomas, vesículas o ulceras dolorosas reaparecen periódicamente. El tratamiento convencional disminuyó su efectividad en cepas resistentes e inmunodeprimidos. Alternativas terapéuticas con extractos de plantas medicinales y potencial antiviral, como Opuntia soehrensii Brito conocida como "ayrampù" en Bolivia, utiliza infusión de sus semillas como analgésico, antidiabético, hipotensor y febrífugo. En vapores por inhalación para afecciones respiratorias; como tintura tópica en lesiones dérmicas de viruela, sarampión y herpes labial. Objetivo. Evaluar la seguridad preclínica de un gel que contiene el extracto hidro-alcohólico de semillas de Opuntia soehrensii en diferentes dosis, aplicado en la mucosa vaginal de ratas Sprague Dawley. Material y métodos. Se ejecutaron protocolos de toxicidad aguda y subaguda para evaluar la respuesta sistémica, a través de marcadores bioquímicos y de comportamiento, y la respuesta local en mucosa vaginal, mediante estudios histopatológicos, en grupos de animales a los que se aplicó el gel con diferentes concentraciones del extracto de Opuntia soehrensii, comparados con un grupo control y otro que recibió solo el vehículo. Resultados. Se encontró que los indicadores sistémicos de comportamiento y ganancia de peso no mostraron diferencias entre grupos. Los indicadores hematológicos y bioquímicos mostraron resultados fisiológicamente esperados y sin cambios en los grupos de estudio. La citología expuso conservación del fenotipo celular para las fases del ciclo estral en todos los grupos. Los indicadores histológicos de reacción local e integridad celular se distribuyeron de igual manera en los todos los grupos. Conclusión. La aplicación de un gel de Opuntia soehrensii no muestra niveles apreciables de toxicidad local y sistémica, lo que permite recomendar la iniciación de estudios de aplicación clínica.

Introduction. Around 3.7 billion people under 50 years of age are infected with HSV-1 and 491 million people between the ages of 15 and 49 are infected with HSV-2 in the world; his symptoms, vesicles or painful ulcers recur periodically. Conventional treatment decreased its effectiveness in resistant and immunosuppressed strains. Therapeutic alternatives with extracts of medicinal plants and antiviral potential, such as Opuntia soehrensii Brito known as "ayrampù" in Bolivia, uses infusion of its seeds as an analgesic, antidiabetic, hypotensive and febrifuge. In vapors by inhalation for respiratory conditions; as a topical tincture in skin lesions of smallpox, measles and cold sores. Objectives . To evaluate the preclinical safety of a gel containing the hydroalcoholic extract of Opuntia soehrensii seeds in different doses, applied to the vaginal mucosa of Sprague Dawley rats. Material and Methods. Acute and sub-acute toxicity protocols were carried out to evaluate local response in the vaginal mucosa, through histo pathological studies, and systemic responses, through biochemical and behavioral markers, in groups of animals to which the gel with different concentrations of the extract of Opuntia soehrensii was applied, compared with a control group and another that received only the vehicle. Results. It was found that the histological indicators of local reaction and cell integrity were equally distributed in all groups. Cytology showed conservation of the cell phenotype for the phases of the estrous cycle in all groups. The systemic indicators of behavior and weight gain did not show differences between groups. Hematological and biochemical indicators showed results ranged in physiologic parameters, without changes in the study groups. Conclusion. The application of a gel from Opuntia soehrensii does not show appreciable levels of local and systemic toxicity, which makes it possible to recommend the initiation of clinical application studies.

In Vivo Acute And Sub-Acute Toxicity Study Of Root Extract Of Carissa Spinarum Linn. In Swiss Albino Mice

Objective: The objective of the present research work had been done to evaluate the toxicity of crude extract of Carissa spinarum in Swiss albino mice. Methods: In studying the toxicity, the Organization for Economic Cooperation and Development (OECD) guidelines were used. Experimental animals (mice), five mice in each, were grouped into four groups; three experimental groups and one negative control. In studying the acute toxicity, 2000, 3000 up to 5000 mg/kg crude plant extract was given orally using standard intragastric oral gavages. For acute toxicity, a single dose was given and gross behavioral changes were recorded. In sub-acute oral toxicity test, Carissa spinarum crude extract was given to the mice by standard intragastric oral gavages at doses of 500, 750 and 1000 mg/kg body weight of hydro-methanolic extract and 200, 600 and 1000 mg/kg of body weight of chloroform extract in every single to 28 d and various hematological and physical parameters were recorded. Results: In acute toxicity, the given dose of the plant extract did not produce significant physical and behavior changes up to the dose of 5000 mg/kg extracts. In addition, no death was occurred in the given doses. In sub-acute toxicity studies of the hydro-methanolic and chloroform extracts, there was no recorded significant change (p>0.05) of hematological and physical parameters in the treated groups when compared to the control groups. Conclusion: from the present study it was revealed that the crude extract of the plant did not produce any significant toxicological effect in the experimental animals and this supports the use of the plant in folk medicines.

Safety and Toxicological Assessment of Allantoin Extract from Cistanches Herba / 中国实验方剂学杂志

Objective: To evaluate the safety of allantoin extract from Cistanches Herba,so as to provide scientific support for subsequent utilization and development of allantoin extract from Cistanches Herba.Method: According to the national standard procedures and methods in food safety and toxicological evaluation,a series of toxicological studies on allantoin extract from Cistanches Herba were conducted,including genetic toxicity and subacute toxicity tests.Result: In the Ames test,with or without mammalina liver microsomal enzymes (S9),allantoin extract from Cistanches Herba in 40,200,1 000,5 000 μg·dish-1 dose range for four bacteria showed no dose-dependent increase.In the micronucleus test,there was no statistically significant difference among each dose group and the negative control group.In the test of mouse sperm aberration,there was no significant difference in the sperm aberration rate among the allantoin extract group and the negative control group.The results of three genotoxicity tests were all negative,indicating no genotoxicity in allantoin extract.The results of 30 days test showed no death and abnormal clinical sign in rats of control group and each dose group (1.100,0.550,0.275 g·kg-1).The body weight,food intake,weekly and total food utilization,weight increment,organ/body ratios,blood biochemical indexes and blood routine indexes had no significant difference among the control group and dose groups.There was no abnormal pathological change in heart,liver,spleen,lung,kidney and testicle of rats in treatment group.Conclusion: Allantoin extract from Cistanches Herba is a non-toxic substance without any genetic toxicity but with a high edible safety.This study provides scientific experimental basis for its safety.

Evaluation of Acute and Sub-acute Oral Toxicity Effect of Aquilaria malaccensis Leaves Aqueous Extract in Male ICR Mice

The study was conducted to investigate the acute and sub-acute toxicity effect of Aquilaria malaccensis leaves aqueous extract (AEAM) towards male ICR mice in terms of body weight, relative organ weight, mortality rate and sperm parameters. In acute toxicity study, a single dose at of 2000 mg/kg was performed. In sub-acute toxicity study, the mice were received normal saline (control group), 50, 100, 150, 200, 500, or 1000 mg/kg of AEAM orally for 21 days of treatment. In sub-acute toxicity study, the number of abnormal sperm were significantly decreased in AEAM 100, 150, 200, 500, and 1000 when compared to the control group. While, the motility of sperm were found to be significantly increased in AEAM 100, 150, 200, and 1000 as compared to the control group. No mortality was recorded in the control group and treated groups in both toxicity studies except for one mouse from AEAM 1000 group. However, the mild sedative effect in terms of the tendency to sleep was clearly noticeable in both toxicity studies. Results indicated that the AEAM can be one of the useful alternative medicine to enhance fertility rate by increasing healthy sperm production.

Toxicity Study of Aqueous Leaves Extract of Jatropha gossypiifolia from Nigerian in Albino Rats: Serum Biochemistry and Histopathological Evaluation

Confounding factors such as plant species, its location and other environmental associated chemistry were reported to influence the dynamics of phytochemicals from being beneficial health-wise to metabolites that facilitate toxic induced effects. This study investigated the oral sub-acute toxicity profile of the aqueous leaf extracts (ALE) of Jatropha gossypiifolia collected from Mubi, Adamawa State, Nigeria. Generalized loss of body weight, weaknesses, dizziness, loss of appetite and restlessness were observed in the acute toxicity study with more severe effects and mortality recorded in the groups exposed to higher doses of 1000 and 2000 mg/kg body weight. In the sub-acute toxicity study, the ALE following the oral administration of 240 mg/kg, 450 mg/kg and 583 mg/kg for 28 days was observed to profoundly alter the normal architecture of the liver and the kidney. The pathological lesions were observed to have disrupted the normal concentration of the serum biomarkers. The ALT concentrations were found to increases to 10.28±1.26 U/L at 250 mg/kg, 9.38±0.57 U/L at 450 mg/kg and 9.31±0.77 U/L at 583 mg/kg respectively when compared to the control (5.86±0.34 U/L). The concentrations of AST were observed to increase to 49.07±16.2 U/L at 250 mg/kg, 47.10±15.42 U/L at 450 mg/kg and 53.07±10.32 U/L at 583 mg/kg respectively when compared to the control (46.11±9.21 U/L). The activity levels of ALP further shows an increase of 379.50±11.31 U/L at 250 mg/kg, 624.90±22.46 U/L at 450 mg/kg and 662.73±28.62 U/L at 583 mg/kg respectively when compared to the control (349.97±71.70 U/L). It is evident from this study that the ALE of the J. gossypiifolia species in addition to its health benefits also contains a cocktail of toxic phytochemicals. Thereby redefined the previous conclusions that the aqueous leaf extract of J. gossypiifolia plant is entirely safe. Thus, reinforcing the importance of the uses of indigenous/site-specific experiment, having in mind that some phytochemicals are sensitive to numbers of confounding factors.

Acute and Sub-acute Toxicity Studies of Ethanol Seed Extract of Raphia hookeri on Swiss Albino Rats.

Aim: To evaluate the acute and sub-acute toxicities of Raphia hookeri (Rh) seed hydroethanolic extract on experimental animals. Materials and Methods: Acute toxicity study was evaluated on Swiss albino mice of both sexes. Administration of a single dose of 4000mg/kg of Rh seed extract by gavages to five mice showed no mortality, hence, its 1/20th dose was used as the highest therapeutic dose. The intra-peritoneal administration produced dose dependent mortality with median lethal dose (LD50) of approximately 323.6mg/kg body weight (bwt). In subacute toxicity study, Wistar rats received daily administration of the extract in the dose range of 50 to 200mg/kgbwt for 30 days. The effects on biochemical, histological and haematological parameters were evaluated. Results: The animals exhibited dose dependent body weight changes. There were some organs weight gains with the exception of the liver and testes which showed comparably lower weight compared to the control. There was a significant (p<0.05) increase in total protein, aspartate aminotransferase (AST) and albumin levels compared to the control while bilirubin and alanine aminotransferase (ALT) levels decreased appreciably at the highest extract dose. The urea level decreased while the creatinine level increased in dose dependent manner. In lipid profile study, total cholesterol, triglycerides and low density lipoprotein cholesterol (LDL-cholesterol) levels showed significant (p<0.05) decrease in value. There was significant (p<0.05) increase in high density lipoprotein cholesterol (HDL-cholesterol). Marked decrease in red blood cells, haemoglobin and haematocrit occurred. The white blood cells also decreased while neutrophil and lymphocytes increased appreciably. The extract caused marked deleterious effect on the testes leading to drastic reduction in sperm cells. Conclusion: The extract caused undesirable effect on the male reproductive organ of the animals making it unsafe for consumption by males of reproductive age.

Sub-acute toxicity of cultured mycelia of Himalayan entomogenous fungus Cordyceps sinensis (Berk.) Sacc. in rats.

Oral administration of laboratory cultured mycelia powder of C. sinensis did not show any sign of toxicity as no significant change was observed in organ weight and serological parameters in rats. However, there was a significant increase in food intake, body weight gain and hematological parameters like WBC, RBC, Hb and lymphocytes in treated groups. Histopathology of vital organs also supported the non toxic effect of C. sinensis. The results conclude that laboratory cultured mycelia powder of C. sinensis is safe and non toxic up to 2g/ kg body weight dose.

Acute and subacute oral toxicity evaluation of benincasa hispida extract in rodents.

The toxicity studies were carried out a 50% aqueous ethanolic extract of Benincasa hispida ( B. hispida )in rodents.The acute toxicity study, B. hispida was found to be well tolerated upto 2000mg/kg, produced neither mortality nor in behavior in mice. In subacute toxicity study, B. hispida at dose level of 200 and 400 mg/kg did not produce any significant difference in their body weight, food and water intake when compared to vehicle treated rats. It also showed no significant alteration in hematological and biochemical parameters in experimental groups of rats apart from a decrease in aspatate transaminase, alanine transaminase and alkaline phosphate content at the dose of 400 mg/kg. Histopathological study revealed normal architecture of kidney and liver of B. hispida treated rats. These results demonstrated that there is a wide margin of safety for the therapeutic use of B. hispida and further corroborated the traditional use of this extract as an anti hepatocarcinogenic agent.

Acute and sub-acute oral toxicity profile of Acorus calamus (Sweet flag) in rodents

Objective: To determine the acute and sub-acute oral toxicity profile of the hydroalcoholic extract of Acorus calamus (HAE-AC) in mice and rats respectively. Methods: In acute toxicity study, mice were assessed to any alteration of general behavior and mortality rate within 24 h. Further, in sub-acute toxicity study, rats were used for assessment of mortality, body weight, hematological, biochemical and histopathological changes. Results: Single oral administrations of the HAE-AC 2500-10000 mg/kg induced increase in general behavioral abnormalities in mice. The mortality rate also increased with increasing dosage (median lethal dose; LD50 = 5 070.59 mg/kg). Daily single oral doses of HAE-AC 200, 500 and 1 000 mg/kg were observed to be well tolerated behaviorally after 28 days of dosing and induced no significant changes in body and organs weights of rats. Further, a mild rise in the levels of alanine transaminase (ALT), aspartate transaminase (AST) and histopatholological changes in liver tissue was noted at 1000 mg/kg dose of HAE-AC. Conclusions: Overall, the findings of this study indicate that, HAE-AC is non-toxic and has at high dose, a mild but acceptable toxicity potential.

Acute and sub-acute toxicity studies of Passiflora nepalensis in rats

This study was designed to elucidate the toxicity of widely used plant Passiflora nepalensis Walp. (Passifloraceae) in rats. We have taken methanolic extract isolated from whole plant of Passiflora nepalensis and studied their toxic effects. Acute, sub-acute toxicities and LD50 values were determined in experimental rats. The external appearance of the dead animals, the appearance of the viscera, heart, lungs, stomach, intestine, liver, kidney, spleen and brain were carefully noted and any apparent and significant features or differences from the normal were recorded after acute treatment with methanolic extract of whole plant of Passiflora nepalensis (MPN). Following the sub-acute administration of MPN for fourteen days, the vital organ such as liver, kidney and heart were carefully evaluated by histopathological and biochemical studies and any apparent and significant changes or differences from the normal were recorded. Pathologically, neither gross abnormalities nor histopathological changes were observed. Oral administration of MPN at the doses of 40, 80, 160 and 320 mg/kg body wt for fourteen consecutive days to male and female rats did not induce any short term toxicity. Collectively, these data demonstrate that the methanolic extract of Passiflora nepalensis have a high margin of safety.

Pharmacognostical standardization and toxicity profile of the methanolic leaf extract of Plectranthus amboinicus (Lour) Spreng.

The present investigation was intended to evaluate the toxicity of the methanolic leaf extract of a traditionally used plant Plectranthus amboinicus (Lour) Spreng. Plant material was analysed for various pharmacognostical parameters as per WHO guidelines procedure i.e., foreign matter, microscopical sections, loss on drying, water and alcoholic extractive values, Total ash, acid soluble ash, heavy metals, phytochemical analysis and toxicity studies. Acute & Sub acute toxicity of the methanolic extract was evaluated in albino mice (Female) after ingestions of the extract during one day (Acute model) and during 28days (sub acute model).The studies on sub acute toxicity reveals that no mortalities or evidence of adverse effects have been observed in Albino mice following acute oral administration at the highest dose of 2000mg/kg crude extracts of Plectranthus amboinicus (Lour) Spreng. Similarly, in sub-acute toxicity study methanolic extract 200,400 mg/kg body wt of Plectranthus amboinicus did not cause any changes in hematological and biochemical parameters. Studies on histopathological examination of vital organs showed normal architecture suggesting no morphological disturbances. Plectranthus amboinicus (Lour) Spreng can be considered as safe as it did not cause either any lethality or adverse changes in the general behavior in mice.

Acute and sub-acute toxicity of an insect pheromone, N-heneicosane and combination with insect growth regulator, diflubenzuron, for establishing no observed adverse effect level (NOAEL).

Aedes aegypti mosquito is one of the most notorious vectors of dangerous diseases like dengue hemorrhagic fever and chikangunya. One method of control of the vectors is by the use of semiochemicals or pheromones. The pheromone n-heneicosane (C21) has been proved to be effective in attracting the female Aedes aegypti to lay eggs in the treated water and the growth of the larva is controlled by insect growth regulator diflubenzuron (DB). This study was planned to assess the safety of C21 alone and the combination with DB. Acute toxicity tests were carried out using two doses, viz., 1600 and 3200 mg/kg and two routes of exposure oral and intra-peritoneal. Dermal toxicity test was carried out in both male and female rats at the dose of 3200 mg/kg. Primary skin irritation test was carried out in rabbits. Sub-acute (90 days) dermal toxicity studies in male and female rats at the dose of 1 and 2 mg/kg via the per-cutaneous route were also studied. Sub-acute (90 days) toxicity test through the oral route was carried out, at doses 125, 250 and 500 mg/kg in male and female rats. The calculated LD50 by ip route and dermal route was more than 5 g/kg in mouse and rats of both the sexes. In the primary skin irritation test no significant changes were noted. In the sub-acute toxicity studies even 500 mg/kg dose was not able to produce toxic response in rats when they were dosed daily for 90 days. The established no observed adverse effect level (NOAEL) was more than 500 mg/kg.