Toxicological and nutritional evaluation of plant cell cultures from scurvy grass (Cochlearia danica) and rowan (Sorbus aucuparia)

Safety evaluation of flexirubin from Chryseobacterium artocarpi CECT 8497: Acute, sub-acute toxicity and mutagenicity studies

Wuzhuyu decoction (WZYD) is a well-known classic traditional Chinese medicine prescription and has been widely used to treat headache, nausea, vomiting, insomnia, etc. However, little published information is available about its safety. Our aim was to investigate the acute and subacute oral toxicity of WZYD extract in rats following the technical guidelines from China’s National Medical Products Administration (NMPA) for single and repeated doses toxicity studies of drugs. Acute oral toxicity was assessed in rats via oral administration of WZYD extract at 4 g/kg three times within a day followed by a 14-day observation period. To evaluate the subacute toxicity, rats were orally administered with WZYD extract at doses of 0, 0.44, 1.33, and 4 g/kg for 28 days. The items examined included clinical signs, body weight, food consumption, hematological and biochemical parameters, bone marrow smear, organ index, and histopathology. After the rats were administered with 12 g/kg (3 × 4 g/kg) WZYD extract, no mortality and toxic effects were observed during the observation period. In the subacute toxicity study, WZYD extract did not cause any significant treatment-related abnormality in each examined item of rats, so the no observed adverse effect level (NOAEL) of WZYD extract for 28 days orally administered to rats is considered to be 4 g/kg, which is approximately 80-fold of its clinical proposed dosage.

Madhulai Manappagu- a well-known sastric and widely prescribed Siddha herbal syrup formulation indicated for treating Veluppu Noi (Anaemia especially Iron deficiency Anaemia) has been in day today practice in Tamil Nadu for a quite longer decades. The syrup is a herbal preparation which has a sweet pleasant odour and a palatable taste, contain the juice of pomegranate (Punica granatum L.) as the main ingredient. Though the formulation is a fruit juice, the safety profile of the syrup is not established and is being marketed without toxicological evaluation. The study is aimed at ascertaining the acute and sub-acute toxicity assessment of Madhulai Manappagu in Wistar Albino rats. The acute and sub-acute (28day repeated oral) toxicity studies were performed as per the guidelines mentioned in the Organization for Economic Cooperation and Development (OECD) 423 (adopted on December 2001) and TG 407 (adopted on October 2008) with slight modifications respectively. For acute toxicity study, three female rats were randomly selected as control; three female rats were randomly selected and were administered a single dose of 5,000mg/kg body weight per oral route. For sub-acute (28day repeated oral) toxicity studies, three doses of test drug MM of 500mg/kg/day (low dose), 750mg/kg/day (intermittent dose) and 1,000mg/kg/day (high dose) were selected for administration. Both sexes of Wistar Albino rats were randomized into four groups of 10 animals each (five males, five females). Group I was kept as control group. Group II, III and IV served as low, intermittent and high doses of MM respectively. Animals were observed for mortality, morbidity, body weight changes, feed and water intake. Haematology, clinical biochemistry, electrolytes, gross pathology, relative organ weight and histopathological examination were performed. In the acute toxicity study, rats showed no toxicological signs on behavior, gross pathology and body weight of rats when treated with a single dose of 5,000mg/kg body weight per oral route. In the subacute (28days repeated oral) toxicity study, rats have showed no significant changes on behavior, gross pathology, body weight, and hematological and biochemical parameters when treated with Madhulai Manappagu in three different doses. The toxicity studies which include both acute and 28days repeated (subacute) oral toxicity studies, revealed no observed adverse effect level (NOAEL) of Madhulai Manappagu in animals. Thus the safety of the drug in human usage was ensured.

This study rigorously evaluated the safety profile of dietary fiber extracted from cassava pulp, a promising functional food ingredient, through acute and 28-day sub-acute oral toxicity assessments in Wistar rats. This research hypothesized that cassava pulp fiber would exhibit minimal toxicity across a range of doses. In the acute study, rats received single oral doses of 175, 550, or 2000 mg/kg, while the sub-acute toxicity study involved daily doses of 250, 500, or 1000 mg/kg, with satellite groups included for reversibility assessment. Comprehensive monitoring encompassed clinical signs, mortality, body weight, food intake, hematological and biochemical parameters, relative organ weights, and detailed histopathological examination. Remarkably, no treatment-related mortality or overt clinical signs of toxicity were observed in either study. The LD50 was higher than 2000 mg/kg for the acute study and the no-observed-adverse-effect level (NOAEL) was determined to be 2000 mg/kg for the acute study and 1000 mg/kg for the sub-acute toxicity study, indicating a high margin of safety. While statistically significant alterations were noted in some hematological, biochemical, and relative organ weight parameters, these changes were not considered toxicologically relevant. Notably, histopathological changes in the lungs were observed across all groups, including controls, warranting further investigation. These findings suggest that cassava pulp fiber is well tolerated at high oral doses, supporting its potential for safe application in food and nutraceutical formulations. However, the observed lung alterations necessitate further research to elucidate their etiology and clinical significance.

Background Alternansucrase is a glucansucrase enzyme that mainly functions in synthesis of glucan-like polysaccharides and can be used for production of novel oligosaccharides and polysaccharides having low glycemic index and prebiotics properties which makes them ideal candidates as dietary fibers and food. From a regulatory perspective, it is necessary to prove the safety of alternansucrase before it can be used in any application as it is a novel enzyme and has not been historically used in food processing. Objectives Alternansucrase obtained from Escherichia coli was subjected to toxicological tests to determine its safety for use in various industrial applications. Design Toxicity studies were conducted at acute oral and repeated sub-chronic (14 days and 90 days) levels in rats following the OECD guidelines. The genotoxicity studies were conducted using the bacterial reverse mutation test as well as in vitro mammalian cell micronucleus test as per the OECD guidelines. Results Alternansucrase did not induce any clinical abnormalities or mortality in rats at the dose level of 2000 mg/kg of body weight in the acute oral toxicity test. A 90-day sub chronic repeated dose oral toxicity study with alternansucrase at and up to dose of 1000 mg TOS/kg of body weight did not show any treatment-related significant toxicological effects on body weight, food consumption, organ weights, hematological and clinical chemistry, or histopathology parameters. Alternansucrase was found to be non-mutagenic up to 5000 µg TOS/plate concentration in the bacterial reverse mutation test. Alternansucrase was determined to be non-clastogenic and non-aneugenic up to the test concentration of 1250 µg TOS/mL during the in vitro mammalian cell micronucleus test. Conclusion The lethal dose (LD50) based on this study is greater than 2000 mg/kg body weight, which falls into the category 5 criteria of Globally Harmonized System (GHS). No-Observed-Adverse-Effect-Level (NOAEL) was concluded to be greater than 1000 mg TOS/kg per day. The studies taken together substantiate the safety of alternansucrase enzyme in various food and associated industries. The present study paves a future for safe use of alternansucrase in varied industrial applications.

Background: 18KHT01 is a novel synergistic composition of Quercus acutissima, Camellia sinensis, Geranium thunbergii, and a small portion of Citrus limon. Our previous report demonstrated that the 18KHT01 exhibits potent antiobesity effects, with synergistic antioxidant, antiadipogenic, and antiobesity activities in diet-induced obese mice. This study explores the toxicity profile and quality control parameters of the 18KHT01 formulation. Methods: Broad-spectrum acute and subacute oral toxicity studies were performed using male and female ICR mice. In order to simultaneous analysis of the 18KHT01 formulation, an ultraperformance liquid chromatography coupled with a diode array detector (UPLC-DAD) method was developed and validated using six marker compounds. Results: Acute oral toxicity evaluation of 18KHT01, administered at single high doses of 2, 2.5, 3, and 5 g/kg, identified 2 g/kg as the no-observed adverse effect level (NOAEL). The LD50 (50% lethal dose) and LD100 (100% lethal dose) of 18KHT01 for male ICR mice were 3.99 and 7.77 g/kg, and those for the female mice were 2.94 and 4.70 g/kg, respectively. In addition, a 30-day repeated dose oral subacute toxicity evaluation indicated that 18KHT01 is safe below 500 mg/kg/day for long-term administration in ICR mice of either sex. UPLC-DAD method validation revealed that each calibration curve for the marker compounds showed good linearity, as well as the validation parameters such as precision, specificity, and accuracy met the acceptance criteria. Conclusion: The present study evidenced the toxicological profile of 18KHT01 polyherbal formulation in mice as well as developed a simple, rapid, and accurate chromatographic method for quality control.

L. sativum L. (family Brassicaceae) is a versatile herbal medicine in Ethiopia. The seed extract is widely employed in traditional medicine, whilst the seed oil is used as edible oil. However, there are no available studies conducted on the safety of the fixed oil of L. sativum seed in Ethiopia. Therefore, this study aimed to evaluate the acute and subacute toxicity of the oil in Wistar albino rats. Acute and subacute toxicity studies were conducted in Wistar albino rats. A single oral dose of L. sativum seed oil was administered, and the animals were followed for 14days. The subacute oral dose toxicity study was conducted in rats of both sexes by repeated 28-day toxicity test as per OECD guidelines. Body weight was measured weekly, and observations of the animals were made regularly throughout the study period. Organ weight, histopathology, hematology, and clinical chemistry data were collected on the 29th day. One-way analysis of variance (ANOVA) was used to compare the means of the comparison groups and the results were presented as mean ± standard deviation, and significance was determined at the P-value of <0.05. In this study, the LD50 of the fixed oil of L. sativum was found to be 2818.32mg/kg. According to the World Health Organization, the oil is classified as slightly hazardous at a single oral dose administration. In the subacute toxicity study, rats treated with the oil showed significant changes behavioral indices such as piloerection, lethargy, and tremor. In addition, gross pathology of organs, body weight, biochemical, and hematological parameters were deranged. The results of the present study demonstrated that the fixed oil of L. sativum has toxic effects. Therefore, it is highly essential to create awareness among the Ethiopian public who use the seeds for medicinal purposes and/or consume the oil as edible oil about the possible health hazards that they may pose.

Mountain ash (Sorbus aucuparia L.) is a tree that is native to northern Europe. It was recently found to be infected with European mountain ash ringspot-associated virus (EMARaV) in Germany and Finland (2,3). EMARaV is not transmitted mechanically and no vector is known, but it is related to negative-sense RNA viruses transmitted by eriophyid mites (3) and represents a new viral species of the genus Emaravirus, which is not assigned to any family. In Finland, EMARaV is common, widely distributed, and detected in all tested ringspot disease-affected and symptomless mountain ash trees (2). Ringspot symptoms occur in mountain ash also in Sweden (west from Finland). In this study, ringspot-affected mountain ash trees were found in Ustreka, which marks the eastern edge of the geologically defined Baltic Shield and the eastern geobotanical borderline of Fennoscandia in eastern Karelia, Russia (1). This border zone can be recognized by changes in vegetation, including occurrence of Siberian larch (Larix sibirica Ledeb.), which does not belong to Fennoscandian flora. Three young mountain ash trees (4 to 5 years old and 2 to 3 m tall) displayed symptoms characteristic of EMARaV at Lake Tsumba (61°50'58″N, 37°45'34″E) in August 2009. EMARaV was detected in collected leaves by reverse transcription-PCR using virus-specific primers as described (2). The nucleocapsid protein genes (944 nucleotides) of the three isolates (Rus1, Rus2, and Rus3) were 97 to 99% identical (GenBank Accession Nos. GU563317, GU563318, and GU563319). On the basis of phylogenetic analysis, the Russian isolates, and three previously characterized isolates from Finland (Ris60, Ris61, and Kuo 12) formed a distinct cluster separate from the remaining 14 previously characterized isolates from Finland and one isolate from Germany. Division of EMARaV to two genetically distinguishable groups was realized for the first time. EMARaV-infected mountain ash trees suffer from chlorosis and growth reduction of varying severity, which impairs their value as an ornamental and for carpentry. To our knowledge, this is the first report in Russia on definite identification of a virus and virus disease in an economically important native plant of northern Europe in the natural habitat as far east as the geobotanical border zone of Fennoscandia.

Mountain ash (Sorbus aucuparia) is widespread in Europe from sea level to the timberline and reaches its most northern range in N-Norway at 71 °N. Sorbus aucupariaoccurs on acid, dry to moist and mesotrophic to oligotrophic soils. The species often grows in secondary forests together with Alnus viridis, Sambucus racemosa, Betula pendula, Frangula alnus and some Salix spp. In most climax forests mountain ash grows only in the shrub layer. In the subalpine belt, the tree is present in green alder scrub and European larch-Swiss stone pine forests. On some N-exposed mountain slopes of the S-Alps, small mountain ash forests with green alder build the upper forest limit above 1500 m. In this study, ecological and phytosociological aspects of this particular forest community along a N-S-transect from the Swiss Central Alps (Grisons) to the South Alps (Ticino/ N-Italy) were investigated. We consider site conditions, local spread, rejuvenation, tree age structure, infestation and phytosociological aspects of S. aucuparia. The occurrence of A. viridis in the N-S-transect and the role of S. aucupariain the secondary and climax forests have been investigated. While in the green alder scrub (Alnetum viridis) of the North single mountain ash trees are present, alder-mountain ash forests have been found at the S-Alpine sites with S. aucupariacovers 25–80%. Here, the Alnetum viridis is absent, and A.viridis is element of the mountain ash forests. In the montane and lower subalpine zone, S. aucupariawill often be replaced later by beech or other climax trees. Above the upper beech limit, we consider the green alder-mountain ash forests as climax forests. At the driest forest sites, grass species are pre-dominant – mainly Calamagrostis spp. and Avenella flexuosa. Based on the earlier provisional phytosociological name of S-Alpine green alder-mountain ash forests, we suggest the name Alno viridi-Sorbetum aucupariae Hari, Leisinger et Zysset 1993 – according to the first description by HARI et al. (1993). Based on our vegetation records of green alder-mountain ash forests, we propose a new sub-association Alno viridi-Sorbetum aucupariae calamagrostietosum subass. nov.

Background: Herbal formulations with antimicrobial and anti-inflammatory properties are commonly used in traditional medicine for oral hygiene. Despite growing popularity, little or no research exists on the safety profile of these products to confirm their long-term safety. An ayurvedic dentifrice formulation was analyzed in Wistar albino rats for acute and subacute oral toxicity, with emphasis on the toxic effects of the product on physiological, hematological, biochemical, and histopathological parameters.Methods: Wistar rats (n = 6) were administered a single dose of the test formulation above 2000 mg/kg body weight and were kept on a 14-day observation period for acute toxicity. Subacute toxicity was induced by daily oral administration of 300, 600, and 1000 mg/kg of the test formulation to male (n = 30) and female (n = 30) rats for 28 days. A 14-day recovery period in the high-dose group was then carried out. Clinical signs, including mortality, food and water intake, body weight, hematological and biochemical parameters, vital organ weights, and histopathological alterations, were assessed.Results: Acute and subacute toxicity studies did not reveal any mortality or significant clinical signs. At 2000 mg/kg body weight, the hepatic changes were minor and reversible. The no-observed-adverse-effect level (NOAEL) was determined to be 1000 mg/kg body weight, as there were no significant changes in biochemical, hematological, and histopathological parameters until this dose.Conclusions: In Wistar albino rats, the tested Ayurvedic dentifrice formulation tapered with quiescent toxicity can be considered safe for oral use. Long-term studies in the form of clinical trials are recommended for human use.

Balanites aegyptiaca (L.) Delile (Zygophyllaceae) is a medicinal plant used in traditional medicine in Burkina Faso to treat helminthiasis. Using an ethnopharmacological approach, the "Institut de Recherche en Sciences de la Santé (IRSS)" has developed a phytomedicine based on an almond extract from the fruit of this plant for the treatment of helminthiasis. This study aimed to assess the acute and subacute oral toxicity and in vivo mutagenicity of phytomedicine. The acute toxicity study was carried out following OECD guideline 423 by administering a single dose of 2000 mg/kg body weight (bw) of phytomedicine orally to female mice. For subacute toxicity, OECD guideline 407 was used. Four batches of 10 rats (5 males and 5 females) were used, including a control batch and three treated batches receiving a daily oral administration of the phytomedicine at doses of 250, 500, or 1000 mg/kg bw for 28 days. The mutagenicity test was performed according to OECD guideline 474; five batches of 10 mice (5 males and 5 females) were given oral doses of 500, 1000 or 2000 mg/kg bw of phytomedicine, colchicine as a positive control at 5mg/kg bw and distilled water as a negative control. In the acute toxicity test, the LD50 was estimated at 5000 mg/kg bw. The subacute toxicity study showed no mortality or signs of toxicity in rats. Biochemical analysis revealed no significant differences between control rats and those treated with the phytomedicine for glucose, creatinine, total cholesterol, triglycerides, AST, chlorine, calcium, and potassium. On the other hand, serum levels of ALT, total protein, PO43-, and Na+ were significantly reduced in some treated rats. The mutagenicity test showed no change in micronucleus frequency between treated and control mice. This study showed that phytomedicine based on an extract of B. aegyptiaca fines would present less danger to users. Keywords: Balanites aegyptiaca, acute toxicity, subacute toxicity, micronucleated erythrocytes, phytomedicine.

Mountain ash (Sorbus aucuparia L.) is a valuable medicinal and food plant that is widely used in practical medicine and in amateur horticulture.&#x0D; The purpose of this study was to determine the content of ascorbic acid, organic acids, tanning substances and flavonoids in the fruits of some representatives of the Sorbus genus of the collection of the South-Ural Botanical Garden-Institute and to identify the most promising taxa. For analysis, fruits were collected in the full ripening phase and dried to air-dry state. Phytochemical studies were carried out according to conventional methods. The object of comparison was the official pharmacopoeia species – Sorbus aucuparia.&#x0D; Analysis of the obtained data found that the fruits × Sorbocotoneaster pozdnijkovii and Sorbus sibirica outperformed the natural form (Sorbus aucuparia) in terms of ascorbic acid content by 11.54 and 43.27%; Organic acids by 24.11 and 60.84%; Tanning substances by 3.49 and 43.23%; Flavonoids by 44.19 and 12.79%, respectively. The obtained results allow to recommend these taxa as promising sources of raw materials for production of biologically active substances in production of natural food products and food additives.

Background: Acute and subacute toxicity screenings of medicinal plants are the fastest way to evaluate the toxicological profiles of medicinal plants. It helps to have a quick idea of the harm or safety potency of drugs. This study aims to conduct an acute and subacute oral toxicity study on the Cyperus Papyrus (CP) ash in Wistar albino rats. CP is traditionally used for managing painful spasms, eye diseases, ulcers, fever, diarrhea, and inflammations. Further study is required to reflect the safety profile of this plant ash. Methods: Acute oral toxicity study of the CP ash was evaluated by using a dose of 2000 mg/kg body weight on rats according to the Organization for Economic Co-operation and Development (OECD) guidelines using the Aot425 software. Subacute oral toxicity was carried out on 4 groups of 6 rats, at doses of 175 mg/kg, 550 mg/kg, 1750 mg/kg, and 1750 mg/kg. The recovery group (control group) received water according to OECD 407 guidelines for consecutive 28 days. At the end of the experiments (day 29), all rats were weighed and sacrificed except the rats of the recovery group which were continued without ash administration for extra 14 days and then weighed and sacrificed. Subsequently, the hematological parameters, plasma biochemical parameters, and histopathological examination were carried out. Results: Upon completion of the acute toxicity, no deaths or signs of acute oral toxicity were observed. Hematologic observations after oral subacute toxicity included a decrease in hematocrit percentage, mean corpuscular volume, and lymphocyte percentage, while there was a slight increase in the mean corpuscular hemoglobin concentration, however, it returned to the normal range in the recovery group. Biochemical tests showed a mild increase in the serum creatine and aspartate aminotransferase levels but they also returned to their normal range in the recovery group. No morphological changes in the kidney, liver, lung, spleen, heart or small intestine were observed after the histopathological examinations. Conclusion: Dried ash of CP is considered to be safe and non-toxic at acute exposure and as the lethal dose 50% value exceeds 2000 mg/kg body weight, it has a mild reversible effect on some hematological and biochemical parameters while no morphological changes of vital organ histopathology occur.

Subacute (28-day) toxicity of furfural in Fischer 344 rats: a comparison of the oral and inhalation route

Введение. В лесопарковой зоне современных урбоэкосистем происходит антропогенная трансформация лесных насаждений, выполняющих средообразующие и средостабилизирующие функции. Нарушенность в них естественных процессов приводит к изменению состава подлесочных видов и вытеснению аборигенных. Цель: мониторинг антропогенной трансформации старовозрастных сосновых насаждений лесопарковой зоны. Задачи: выявление ценотической роли Sorbus aucuparia L., определение её экологических ниш и плотности ценопопуляций. Объекты исследований: ценопопуляции Sorbus aucuparia L. пяти лесопарков Екатеринбурга: им. Лесоводов России, Нижнеисетский, Шарташский, Шувакишский и Юго-Западный. Методика работы заключалась в маршрутном обследовании лесопарков, расположенных на разном удалении от города, закладке 16 пробных площадей для установления плотности (шт./га) ценопопуляций Sorbus aucuparia L. в зависимости от полноты древостоя. Установление экологических ниш в перестойных сосновых насаждениях лесопарковой зоны Екатеринбурга проведено с помощью программы АРМ «ЛесФонд». Результаты. Доминирующий подлесочный вид Sorbus aucuparia L. представлен фрагментами поселений разных возрастных групп в 12 типах леса. Его максимальные площади установлены в сосняках ягодниковых, в удалённых от города лесопарках, Шувакишском и Нижнеисетском. Устойчивость ценопопуляций Sorbus aucuparia L. подтверждается их высокой плотностью и многообразием экологических ниш. Оптимальная экологическая ниша Sorbus aucuparia L. в перестойных сосновых насаждениях – сосняк ягодниковый и сосняк разнотравный. Наибольшая плотность ценопопуляций Sorbus aucuparia L., как показателя освоенности экологической ниши, определена в сосняке ягодниковом Шарташского лесопарка с полнотой соснового древостоя 0,7. Основным конкурентом Sorbus aucuparia L. в захвате подлесочного пространства исследованных лесопарков является Malus baccata (L.). Выводы. Древостой сосны обыкновенной во всех лесопарках является фитоценотической защитой для аборигенных подлесочных видов от инвазионных ценопопуляций Malus baccata L. и других древесных интродуцентов. Introduction. In forest parks of modern cities, an anthropogenic transformation of forest plantations, performing habitat forming and environment regulation functions, is observed. The break of natural processes leads to the change in the composition of undergrowth species and supplantation of native species. Thegoal of this research is to monitor the anthropogenic transformation of old-aged pine stands in the urban wooodlands. Tasks: to reveal the cenotic role of European mountain ash (Sorbus aucuparia L.), define its ecological niches and cenopopulation density. The cenopopulations of Sorbus aucuparia L. of five forest parks of Ekaterinburg are the object of the research. These parks are the forest park named after Foresters of Russia, Nizhneisetskiy forest park, Shartashskiy forest park, Shuvakishskiy forest park, and Yugo-Zapadnyy forest park. The methodologyincluded a route survey of forest parks located at different distances from the city and establishment of 16 study plots in order to determine the density (pcs/ha) of Sorbus aucuparia L. cenopopulations depending on the normality. Identification of ecological niches in the overmature pine plantations of Ekaterinburg forest-park zone was carried out under the “LesFond” program. Results. The dominant undergrowth species of Sorbus aucuparia L. is represented by fragments of populations of different age groups in 12 forest types. It takes the maximum areas in berry pine forests in remote from the city forest parks (Shuvakishskiy and Nizhneisetskiy forest parks). The stability of Sorbus aucuparia L. cenopopulations is confirmed by their high density and variety of ecological niches. The optimal ecological niche of Sorbus aucuparia L. in the overmature pine plantations is a berry pine forest and a herb pine forest. The highest density of Sorbus aucuparia L. cenopopulations, as an indicator of the development of an ecological niche, was found in the berry pine forest of Shartashskiy forest park with 0.7 density of pine. Malus baccata (L.) is a chief competitor of Sorbus aucuparia L. in capturing the undergrowth area of the studied forest parks. Conclusion.Scots pine stand is a phytocenotic protection for native undergrowth species from invasive cenopopulations of Malus baccata L. and other woody introduced species.