Decrease in the volume of plasma water (hyperproteinemia) is noted in cases of dehydration due to inadequate water intake or excessive water loss, as the case in severe vomiting or diarrhea [43]. The initial mean body weight of control group was 144.70 21.89 g, and final mean body weight was 203.22 31.07 g. The initial mean body weight of rats treated with the dose of 500 mg/kg was 148.48 11.94 g, and final mean body weight was 206.90 17.19 g. The initial mean body weight of rats treated with the dose of 1500 mg/kg was 139.80 9.26 g, and the final mean body weight was 193.48 16.63 g. Bar graph of mean body weight change in rats treated with 500mg/kg and 1500mg/kg of 80% methanol extract as compared to the control group during subacute toxicity study. dc. In contrast to this, the study conducted in Nigeria on antivenom studies of S. guineense extracts against Naja katiensis venom rats revealed that the methanol leaf extract has produced no significant change in blood glucose level [42]. Clinical observation was carried out for 28 days and their weight was measured weekly for four weeks. Group III served as control group and received 80% methanol, the vehicle to dissolve the extract. In the present study, the mean values of urea have been shown with a slight increment at dose of 500 and 1500 mg/kg even though not significant and this was not associated with the histopathological changes of the kidney. Subacute toxicity study examines toxicity caused by repeated dosing over an extended period of 28 days of oral administration in rodents. So, sections were placed in xylene I for 5 minutes and xylene II for 2 minutes again to remove the paraffin from tissue and hydrated with decreasing concentrations of absolute I, II and 95% alcohol for two minutes each, 70% of alcohol for three minutes, and 50% alcohol for five minutes. Ghildyal P., Groslash T. E., Skogsrud M., et al. After sacrificing the rats, gross pathological observation was carried out on vital organs. In the present subacute toxicity study, the rats that were treated with 80% methanol extract of leaves at doses 500 mg/kg and 1500 mg/kg showed no signs of morbidity and mortality. They were housed in standard cages and kept under standard condition at a temperature of 22C ( 3C), with 12hrs light/12hrs dark cycle [30]. This was supported by the absence of histopathological changes in the liver of treated rats. The 80% methanol extract of the leaves was then administered orally at the doses of 2000 mg/kg (group I) and 5000 mg/kg (group II) body weight of rats in the test groups. The acute toxicity study of the 80% methanol extract of the S. guineense leaves did not produce adverse effects on the behavior and gross pathology of the rats at treated doses. In: Hodgson E., Levi P. E., editors. Creatinine is produced endogenously and released in to body fluids at a constant rate and its plasma concentration is maintained predominantly by glomerular filtration. Almost any substance can be harmful at some doses but, at the same time, can be without harmful effect at some lower dose. However, the weight gain difference between control and treatment groups was statistically insignificant. The control group received distilled water, while the experimental groups received 500 mg/kg and 1500 mg/kg 80% methanol extract of S. guineense leaves orally for 28 days. HHS Vulnerability Disclosure, Help Another study by Nigatu [32] showed that the LD50 (p.o.) There were no signs of toxicity observed in the kidney and liver sections of treated rats. The mean serum glucose level showed significant decrease (p < 0.05) at both doses compared with the controls. Body weight change is an important index for assessment of toxicity [33]. Its watery extract of fruits and bark has activity against different strains of bacteria responsible for diarrhea [22, 23]. The animals were acclimated to laboratory conditions for 5 days. In the acute toxicity study, rats treated with 2000 mg/kg and 5000 mg/kg showed no toxicological signs observed on behavior, gross pathology, and body weight of rats. Following the fasting period, all animals were weighed, and the doses were calculated based on their body weight. After examination, photomicrographs of selected samples of liver and kidney section from both the treated and control rats were taken under a magnification of x20 objective using (EVOS XL, USA) automated built-in digital photo camera. Proceedings of the 9th World rabbit congress; 2008; Verona, Italy. Plant medicine is the oldest form of health care known to mankind. No morbidity or mortality was observed in the treated groups at both doses during acute toxicity study. In this study, the mean values of AST and ALT at dose 500 mg/kg were high whereas at dose 1500mg/kg decreased compared with control, but it was not significant. When a cell is damaged, it leaks this enzyme into the blood. Each animal was assigned a unique identification number. Tadesse M., Demissew S. Medicinal Ethiopian plants: inventory, identification and classification. In the treated rats of kidney sections revealed normal glomerulus (G), Bowman's capsule lined with outer parietal layer/squamous cells (SC) and inner visceral layer/podocytes (P), urinary space (US), proximal convoluted tubules (PCTs) lined by simple cuboidal epithelium with brush border, distal convoluted tubules (DCTs) lined by simple cuboidal epithelium with more nuclei per cross-section, and macula densa (MD) with taller cells around the vascular pole. The values in the extract treated rats were not significantly different from the control (Table 4). Canning Vale WA, ANCCART News 1993; 6(4): 1-4. will also be available for a limited time. The twigs and leaves have anthelmintic (hookworm). In this study the measurement was within reference value and was supported by the absence of histopathological changes of the kidney. Mukherjee P. K., Saha K., Murugesan T., Mandal S. C., Pal M., Saha B. P. Screening of anti-diarrhoeal profile of some plant extracts of a specific region of West Bengal, India. Abou-Mansour E., Djoukeng J. D., Tabacchi R., et al. The authors presented in the thesis were Million Loha Lorato, Abay Mulu, Solomon Mequanent, Wondosen Ergetie, and Bekesho Geleta. Pieme C., Ngoupayo J., Nkoulou C., et al. The gross pathological examination of the liver and kidneys of the treated rats showed no change in color, shape, size, and texture compared to the control group. The outcome of this study may fill the gap of the previous studies on S. guineense and also it is hoped to provide some additional evidence for recommending further studies to assess toxicity profiles associated with the use of herbal preparations of this plant. The difference between groups with respect to variables under investigation was significant at P value of less than 0.05. CV= central vein, EC= endothelial cells, H= hepatocytes, KC= Kupffer cells, S= sinusoids, BD= bile duct, HA= hepatic artery, and PV= portal vein. Malele R. S., Moshi M. J., Achola K. J., Munenge R. W. Pharmacological properties of extracts from the stem bark of Syzygium Guineense on ileum and heart of laboratory animals. Finally, the tissues were embedded in paraffin wax in square metal plates forming tissue blocks, whereby each tissue block was labeled and stored at room temperature till sectioned. Loha M. Acute and subacute toxicity of hydroethanolic extract of syzygium guineense leaves on the histology of the liver and kidney and biochemical compositions of blood in rats. Increment in body weight determines the positive health status of the animals [37]. Ezenyi I. C., Mbamalu O. N., Balogun L., et al. Values are expressed as mean SEM, N= 4/group. In addition, there was no significant difference in the absolute weight of liver and kidneys of treated rats compared to control group. In the histopathological study of the kidney, rats treated with both doses (500 and 1500mg/kg) of the extract showed no significant difference compared to controls. After administering the plant materials, the animals were kept under close observation continuously for 1 hour and intermittently for 4 hours and thereafter once every 24 hours for the next 14 days. OBN6.34/2008) through Ethiopian Public Health Institute (EPHI). Many medicines commonly used today are of herbal origin. The supernatant portion was filtered with 0.1 mm2 mesh gauze and then with Whatman grade 1 filter paper with pore size of 11m. At the end of the experiment, blood samples were collected for hematology and clinical chemistry evaluations. Chambers F. L. A textbook of modern toxicology. Antidiabetic potentials of Syzygium guineense methanol leaf extract. The sections were dehydrated with increasing alcohol concentration of 50%, 70%, 95%, absolute I and II for two minutes each. These remedies, with a considerable extent of effectiveness, are socially accepted and economically viable and, mostly, are the only available source [4, 5]. The studies also indicated that the methanol extract of the bark has a hypotensive effect [14], antihypertensive and vasodilatory effect [15], analgesic and anti-inflammatory effect [16], antimalarial activity [17], and antidiabetic activity [18]. In the current histopathological examination of the liver, rats treated with doses of 500 mg/kg and 1500 mg/kg of the 80% methanol extract of the leaves of S. guineense showed no change in the microscopic structure of the liver. The large array of toxic chemicals produced by plants (phytotoxins), usually referred to as secondary plant compounds, is often held to have evolved as defense mechanisms against herbivorous animals, particularly insects and mammals. The present finding from the acute toxicity study agreed with the study reported by other researchers that there was nontoxic nature of the aqueous leaf extract of S. guineense up to 6000 mg/kg [18]. Antibacterial triterpenes from Syzygium guineense (Myrtaceae). Bethesda, MD 20894, Web Policies Rahman M. F., Siddiqui M. K., Jamil K. Effects of vepacide (azadirachta indica) on aspartate and alanine aminotransferase profile in a subchronic study with rats. Blood samples in the test tubes without anticoagulant could clot and sera were obtained by centrifuging the blood using an electrical centrifuge (HUMAX-K, HUMAN-Germany) from which blood chemistry (glucose, urea, creatinine, total protein, ALT, and AST) was studied to test renal and hepatic functions. The rats were then anesthetized under diethyl ether. The discrepancy between our findings with Amare's report might be because of the duration of exposure difference (42 days versus 28 days), species difference (mice versus rats), and the extracting solvent used (waters versus 80% methanol). Federal government websites often end in .gov or .mil. Traditional medicine, African health monitor. The current result disagreed with findings of study done on oral treatment of the aqueous leaf extract at dose of 200, 400, and 600mg/kg for six weeks in mice at which two mice died at days 32 and 40 from groups treated with 600 and 200mg/kg body weight of the extract, respectively [35]. Histopathological studies of the kidneys sections of rats treated with doses of 500 mg/kg (Figures 7(a) and 7(b)) and 1500 mg/kg (Figures 8(c) and 8(d)) showed no significant microscopic changes compared with the controls (Figures 6(a) and 6(b)). Photograph of liver and kidneys from the subacute toxicity study. Abay Mulu, Solomon M. Abay, and Bekesho Geleta helped to design the study and supervised the study. The result agreed with the study conducted on aqueous extract in normoglycemic and diabetic mice [18]. The mean absolute weights of the liver were 5.98 0.82 g (at 2000mg/kg), 5.03 0.55g (at 5000mg/kg) compared with the control (5.40 1.81) g. The mean absolute weights of the kidneys were 1.10 0.18 g (at 2000mg/kg) and 1.15 0.16 g (at 5000mg/kg) compared with the control (1.15 0.54) g (Table 1). Herbal medicine flourishes today as the primary form of medicine for perhaps as much as 80% of the world's population [1]. Meanwhile, subacute toxicity study of the 80% methanol extract of the S. guineense leaves did not adversely affect the body weight and hematological and biochemical parameters of tested doses. Vahalia M. K., Thakur K. S., Nadkarni S., Sangle V. D. Chronic toxicity study for tamra bhasma (a generic ayurvedic mineral formulation) in laboratory animals. In the present study, all biochemical parameters did not show significant changes except mean values of serum glucose. Learn more (c) and (d) Kidney sections of control rats (H&E-x400). Before In the subacute toxicity study, the biochemical parameters (except glucose) of the treated groups (500 mg/kg and 1500 mg/kg) were within the reference range for rats and were not significantly different from the control group. Marcela E., Silva B., Jaime C., Pilar P. Effects of solanum nudum extraction on the liver of mice infected with plasmodium berghi. The liver and kidney sections taken randomly for tissue processing were fixed in 10% neutral buffered formalin (NBF) overnight at room temperature. This was inconsistent with the finding of Amare [35], who reported a significant increase in the liver weight ratio in mice treated with 200 mg/kg and a significant increase in the right kidney weight ratio in mice treated with 600 mg/kg. Therefore, the oral LD50 of the 80% methanol extract of the leaf of S. guineense was greater than 5000mg/kg. The result was further supported by the values of biochemical parameters of the blood (such as urea, creatinine, and total protein), which are main indicator of kidney damage [47]. The authors are thankful for the financial support provided by School of Graduate Studies of Addis Ababa University (AAU) and Ministry of Finance and Economic Development (Grant no. In most developing countries, the indigenous modes of herbal treatment are parts of the culture and the dominant method of healing therapy. (a) and (b) Photomicrographs of liver section of control rats (H and E, X400). The The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as potential conflicts of interest. Accessibility In many countries, herbal medicines and related products are introduced into the market without safety or toxicological evaluation. toxicity Despite these all efficacy studies on various parts of the S. guineense, there is a limited safety study conducted for this medicinal plant. Pass D., Freeth G. The rat. Satyapal U. S., Kadam V. J., Ghosh R. Hepatoprotective activity of livobond a polyherbal formulation against CCl4 induced hepatotoxicity in rats. The Ethiopian Flora is estimated to consist of between 6000 and 7000 species distributed in about 245 plant families. This finding agreed with work of Amare [35] who reported that mice treated at a dose of 200 and 400 mg/kg body weight of the aqueous leaf extract of S. guineense showed no histopathological changes compared to control group, whereas the tissue morphology of mice treated with 600 mg/kg showed hemorrhagic centrilobular necrosis and fatty cytoplasmic vacuolation of the hepatocytes [35]. The mean body weight gain for the control rats was 19.10 g. The mean body weight gain for rats treated with 2000 mg/kg and 5000 mg/kg was 18.9 g and 17.8 g, respectively. Traditional herbal drugs of southern Uganda. The .gov means its official. Finally, on the 15th day, their final weights were measured, and gross physical examinations were carried out. Syzyguim guineense extracts show antioxidant activities and beneficial activities on oxidative stress induced by ferric chloride in the liver homogenate. CV= central vein, EC= endothelial cells, H= hepatocytes, KC= Kupffer cells, BD= bile duct, HA= hepatic artery, and PV= portal vein. The healthy and nonpregnant Wistar rats of both sex with age of 8 to 10 weeks and above and weight of 120g140 g were obtained from Physiology Department, Addis Ababa University. Values in the sera were analyzed using Automated Clinical Chemistry Analyzer (AUTO LAB 18, clinical chemistry analyzer, Italy). The plant materials were cleaned of extraneous materials and 80% methanol extract of S. guineense was prepared for the study. Therefore, the LD50 of the extract could be greater than 5000 mg/kg. But there were some changes in histology of kidney. During the subacute toxicity study, the treated rats showed gradual normal increase in their body weight. The liver and kidneys have fundamental roles in the metabolism and excretion of drugs or plant products. Plant medicine is the oldest form of health care known to mankind. The mean body weight gain for rats treated with 2000mg/kg, and 5000mg/kg was 18.9 g and 17.8 g, respectively. In the present subacute toxicity study, the hematological parameters (RBC, WBC, PLT, HGB, HCT, MCV, MCH, and MCHC) were within the reference range for rats. All groups of the rats fasted overnight prior to administration. Acute toxicity test assesses the adverse effects that occur within a short timeafter administration of a single dose of a test substance. Evidence-based Complementary and Alternative Medicine : eCAM, http://www.wpro.who.int/health_technology/book_who_traditional_medicine_strategy_2002_2005.pdf, http://www.jaicaf.or.jp/publications/ethiopia_ac.pdf, http://www.kafa-biosphere.com/assets/content-documents/KafaFloral-Survey-Final-Report.pdf, http://etd.aau.edu.et/handle/123456789/2809, http://iccvam.niehs.nih.gov/SuppDocs/FedDocs/OECD/OECDtg425.pdf, http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.626.7531&rep=rep1&type=pdf, http://etd.aau.edu.et/handle/123456789/508, Mean corpuscular haemoglobin concentration, Organization of Economic Cooperation and Development. No mortality was observed in the treated groups, i.e., at 2000 mg/kg and 5000 mg/kg during the study period. Djoukeng J. D., Abou-Mansour E., Tabacchi R., Tapondjou A. L., Bouda H., Lontsi D. Antibacterial triterpenes from Syzygium guineense (Myrtaceae). The dehydrated tissues were cleared in two changes of xylene-for one and half hours and two and half hours. During the subacute experimental period all groups of rats showed gradual and normal increase in their body weight (Figure 2). The mean values of total protein were within the reference range for rats, which was also supported by the absence of histopathological changes in the kidneys of treated rats. Its bark is used in traditional medicine to treat gastointestinal upsets and diarrhea [1921]. In the present study, the changes in hematological parameters of treated groups were not statistically significant compared with control group. Effect of 80% methanol extract of leaves of S. guineense on hematological parameters. Eighteen rats were randomly distributed into three groups (I, II, and III) each consisting of six rats (three female and three male) per group. It is widely distributed in sub-Saharan Africa [9, 10]. The abnormal elevation of the liver enzymes (ALT and AST) is usually associated with liver damage or alteration in bile flow. Although the exact number is still unknown, many the species, i.e., about one-third of the families, have been employed in traditional medicinal practices [7]. Serum total protein change is caused by a change in the volume of plasma water and a change in the concentration of one or more specific proteins in the plasma.