In the present study, acute oral toxicity study of A. album(6C, 30C, 200C) was conducted in Wistar albino rats as per the OECD 423, 2002 guideline with slight modifications. It was used to estimate the LD50 range according to GHS classification. A. album(6C, 30C, 200C) administered at a dose of 2000 µl/kg on day 1 and animals were observed for the period of 14 days. During the experiment, mild lethargy, and dehydration was noticed up to 24 hours of treatment of A. album (6C, 30C, 200C) and alcohol (90%). However these symptoms disappeared after 24 hrs and all the animals were recovered. Moderate abdominal breathing, vocalization and dyspnoea were noticed from 30–40 minutes to 4 hour in some animals which worsens in 24hrs in females and lead to death of animals at 2000µl/kg of A. album (6C) and Alcohol (90%). However, clinical signs and mortality observed is not related with the drug as these symptoms and mortality was visible in vehicle control group also suggested that the observed clinical signs and mortality is due to high concentration of alcohol i.e. 90% dispensing alcohol. There was no drug related mortality observed in animals after the treatment of A. album in its potencies in both the step.
Body weight, percentage body weight changes (change in body weight with respect to Day 01) for all the animals in either step did not reveal any treatment related effect throughout the experiment when evaluated statistically. The gross necropsy examination conducted on day 15 of step 01 and step 02 revealed no abnormal changes in either external surface of the body or internal organs observed. In the present investigation, the animals were administered ten times the dose used clinically i.e. 2000 µl/kg bodyweight for investigating the toxic manifestations due to the over dosage of the test substance within 24 hours. As a result of an implicit assumption, we concluded that the clinical symptoms and mortality observed could be attributed to the vehicle (alcohol) rather than the medicine itself. It is so because the clinical signs and mortality were commonly witnessed in vehicle control (alcohol) as well as in all the treatment groups (6C, 30C, 200C). It is reasonable to conclude that there were no toxic signs or symptoms as a result of the greater dose, which is reinforced by the fact that no mortality was observed due to treatment during the experiment.
Repeated dose 28-day oral toxicity study was performed in Wistar albino rats in accordance with OECD 407, 2008 with slight modifications to investigate the repeated treatment effect of A. album on potential targets of toxicity. In present study we evaluated the effect of A. album (6C, 30C, and 200C) 28 day repeated exposure on Body weight, feed consumption, hematology, biochemistry and organ histology. There are no treatment related changes in behavioral and clinical signs such as gait, fur color, eating, sleeping secretions excretions and autonomic activity were found during the experiment. The body weight change is a sensitive marker of overall health of animal. According to OECD and CCAC guidelines more than 20% loss of body weight considered as one of human end point of study (Olfert et al., 1998; Chitra et al., 2015). The body weight change depends on feed consumption, feed palatability, feed nutritional content, digestion, absorption (Wren et al., 2002). The body weight, relative body weight and feed consumption recorded on day 1st, 7th ,14th ,21st and 28th found to be consistent throughout the study in A. album treated groups and normal control.
Hematological parameters are important diagnostic tools in clinical setting to evaluate the toxic effect of chemical and herbal substances (Rao, 2006; Yakubu et al., 2007). Hematological parameters play pivotal role in monitoring functioning of bone marrow and intravascular system, because bone marrow is highly sensitive to toxic substances (Mukinda et al., 2007). Moreover, studies have shown that hematological alterations in rodents found to be predictive of toxicity in humans as supported by data extrapolated from animal studies (Olson et al., 2000). In our study, we found no treatment related changes in the hematological parameters compared to normal control animals. The non-significant changes in blood parameters suggest that A. album (6C, 30C, 200C) is non-toxic to hematological parameters.
In toxicological studies, clinical biochemistry plays major role in assessing toxic potential of the test substance. Serum biochemical parameters (Blood glucose, total proteins, Serum creatinine, ALP, SGOT, SGPT, Urea, BUN, cholesterol, HDL, LDL) are important in evaluating functioning of liver and kidney as they are essential organ for survival, metabolizing and detoxifying chemicals and drugs (Olorunnisola et al., 2012; Ekanayake et al., 2019). The liver is the main organ involved in drug metabolism, cholesterol synthesis and degradation. In addition, it have role in glucose synthesis and generate glucose from hepatic glycogen stores (Anderson et al., 2008). In the present study, results indicate that A. album treatment has no effect on lipid and carbohydrate metabolism. Serum ALP, SGOT, SGPT assessed in toxicity studies to track liver function. Elevated levels of ALP, SGPT and SGOT are indicator of hepatic injury, cirrhosis and myocardial infarction [25], (Schalm, 2000; El Hilaly et al., 2004). The levels of ALP, SGPT and SGOT in our study found to be similar in A. album treated and control is group animals illustrating that A. album is not causing any liver toxicity. Total bilirubin an indicator of liver function and cholestasis is basically a byproduct of heam catabolism. It is found reduced in conditions such as aplastic anemia (Loge et al., 1958; Rubin et al., 1968; Vítek, 2012). .During the study levels of total bilirubin and BUN are not significantly different in arsenic treatment and control group. Studies have shown that BUN is a diagnostic marker for assessing renal function in clinic settings (Wallig et al., 2017). Moreover, the level of urea and creatinine which are indicators of glomerular filtration rate for assessing the renal function found to be non-significant in arsenic treated groups compared with normal control. These results imply that A. album (6C, 30C, 200C) has no influence on renal function.
Toxic substance cause an array of metabolic reactions in vital organs (brain, lung, liver, stomach, kidney, kidney, thymus, spleen, heart, testis and ovary)resulting in change in color and architecture, hypertrophy, edema some of the initial indications of toxicity (Nigatu et al., 2017). Macroscopic examination of vital organs during necropsy of A. album treated animals in both acute and sub-acute toxicity studies did not show any lesion, hypertrophy, change in color and texture change, compared with normal control. In addition, organ weight a significant indicator of physiological and pathological condition of animals. Absolute organ weight and relative organ weight are fundamental markers determining any treatment related injury in internal organs (Michael et al., 2007). Since, there was no significant change in absolute and relative organ weight of internal organs of treatment groups was observed in the study compared with control group animals. Hence, A. album treatment neither interfered with physiology nor produced pathological change in animals.