In Vitro Screening for Antitrypanosomal Potentials of Punica granatum L. Leaves Crude Extracts

  1. Muhammad Muhsin Fathuddin
  2. Helen Ileigo Inabo

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Abstract

Punica granatum L. leaves were examined for potential antitrypanosomal properties. These leaves were acquired and identified at Ahmadu Bello University (ABU) Herbarium Unit in Zaria. Following drying, the following solvents—chloroform, ethyl acetate, and ethanol—were used for exhaustive Soxhlet extraction. The crude extracts phytochemical analysis and in vitro antitrypanosomal capability was performed on Trypanosoma brucei brucei, obtained from The National Research Institute for Chemical Technology (NARICT), Basawa, Zaria. In 96-round-bottom-well microtiter plates, the in vitro trypanocide activity was evaluated in duplicate. Wet and thick blood film and rapid matching were used to examine the antitrypanosomal activity under 400x magnification. The antitrypanosomal activity was determined using the chloroform, ethanol and ethyl acetate extracts ranging from 006.25 to 400.00 mg/ml. The red blood cells (RBCs) were lysed at all concentrations between 200.00 and 400.00 mg/ml, while between 006.25 and 100.00 mg/ml, the RBCs were relatively intact. It was observed that as concentrations declined, the parasite's motility decreased. The parasite's motility entirely stopped after 60 minutes, whereas it continued for an additional 80 minutes in the negative control. A standard drug that was made per the manufacturer's instructions and used as the positive control which cleared everything in less than a minute. Accordingly, the chloroform extract despite lysing the RBC, did not affect the parasite and did not kill it. At a minimum concentration of 6.25 mg/ml, the Punica granatum L. ethyl acetate and ethanol extract can potentially operate as antitrypanosomal agents. This research may help in the development of novel antitrypanosomal medications from Punica granatum L.

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  1. Access Microbiology

    The present paper is not suitable for publication in Access Microbiology due to the major reasons listed below: 1. The data is too preliminary. The phytochemical characyerisation of the leaf extracts is overly simplistic, and a more thorough caracterisation will be necessary (the isolation of different fractions and their chemical characterisation would be of particular interest). The determination of the antitrypanosome activity of these extracts is solely based on the motility (and not necessarily the survival) of trypomastigote forms; assays such as MTT and Alamar Blue are preferred. 2. The concentrations of DMSO utilised are high and important controls are not shown. Controls containing only DMSO at the same contrations as utilised for the leaf extracts are not shown. This type of control is paramount as 10% DMSO (as in the highest concentrations tested) are known to rapidly kill trypanosomes. Moreover, data on the lysis of red blood cells was not shown, but only mentioned in the text. As a suggestion, the authors should include symbols in the graphs, as some colours shown are very similar to each other, which compromises data interpretation. 3. The text is overly simple. It lacks methodological details and poorly describes and discusses the results. 4. Technical language is incorrect. For instance, in many cases, names of species are not highlighted. Also, 40 grams correspond to 40,000 mg. This paper may be considered for resubmission should the relevant points above are adderessed.

  2. Version published to 10.1099/acmi.0.000875.v1 on Access Microbiology