3.1 Result
Concentrations (100, 250, 500) PPm of the nano composite of Citrullus colocynthis were used to demonstrate their effect on T. vaginalis compared to non-treatment (control) as shown in Table (1), Figure(1) that indicate a reduction in the numbers of parasite in an inverse relationship with the increase in the different concentrations in the solution. When analyzing the results statistically, it was found that there are significant differences at a probability level p ≤ 0.05 between the growth rate of treated and untreated samples, as it had a clear toxic effect on growth. The extract concentration that caused inactivation of 50% of parasites within 24 hours of growth (IC50) was 99 ppm Table(3). It was also observed that the percentage of growth inhibition increased from 17.8% to 46.1% when increasing concentrations compared to an inhibition rate ( 43.4%) when using metronidazole within 12 hours of growth. While the rate of inhibition increased from 43.7% to 69.1% within 24 hours, while metronidazole inhibited growth was 70.4% at the same time. After 72 hours of growth, the inhibition rate increased from 53.4% to .89.8% compared to the drug's inhibition rate of 87%.
Table (2) indicated the effect of concentrations (100, 250, 500) ppm of the nano composite For Capparis spinosa fruits on the number of treated parasites compared to untreated. As the increase in concentration led to a decrease in the number of parasites, with an inverse relationship during different growth periods. It was clear that the IC50 concentration of C. spinosa fruit extract was 99.4 ppm during the logarithmic phase compared with the control Table (3), as this concentration was used in the subsequent experiments. The results of the statistical analysis showed that there are significant differences at the probability level p ≤ 0.05 between the rates of the numbers of parasites treated with the concentrations of this extract and the rates of the numbers of untreated (control) during the different growth periods.
Table (2) and Figure(2) indicated that the percentage of inhibition increased from 16.9% to 43.1% when increase the concentrations within 12 hours of growth, while metronidazole had a higher effect (43.4%). The rate of inhibition of metronidazole was higher after 24 hours of growth, reaching 70.4%, while the rate of inhibition using concentrations of plant extract increased from 42.5% to 67.4%. After 72 hours of growth, the highest inhibition rate was recorded by the nanoscale solution, and it was 87%, which is the same as that of metronidazole.
The extracts used in the experiment showed a weak ability to hemolysis red blood cells. The highest effect of C. spinose and C. colocynthis was at a concentration of 500 ppm, with a percentage of degradation (2.8%) and (2. 3%), respectively, while the plants showed the lowest percentage of degradation at 100 ppm when it was recorded (0.3%) and (0.1%), respectively figure (3).
3.2 Discussion
The present study indicated the ability of nano composites of C. colocynthis to remove T. vaginalis in a more effective concentrations than metronidazole, and gradually upon increasing concentration. C. spinose was equally effective as metronidazole. On the other hand, both plants showed a slightly toxic in hemolysis.
Metronidazole breaks down DNA by releasing free nitrogen radicals and then the cell will die, as the parasite metabolism depends on the generation of ATP through oxidation of acetate produced by pyruvate and in the presence of Co A, nitrogen in the drug works to capturing an electrons that produced by the transport enzymes chain, thus forming free nitrogen radicals and the cell dies 15. However, many strains of this parasite existed that were resistant to 5-metronidazole and its derivatives16 due to the frequent use of this drug 17 or to genetic changes in the parasite 18.
Medicinal plants have been used throughout a person's life and have been accompanying his life path since existence on the earth. It is known that many medicinal plants possess many bio-active ingredients such as phenolic, alkaloids, tannins, terpenes, steroids, saponins, flavonoid compounds11, but the large size of these particles made it difficult to pass through the lipids of the plasma membrane, thus their effectiveness was weak, and an effective effect could only be obtained with an increase in the concentration of these substances, which causes toxicity to living cells and produces negative effects19. Preparation of nanocomposites from these plants solves the large size problem and improves their ability to pass through the plasma membrane by generating nanocomposites surrounded or carried by oil 20. Also, these compounds become more stable and soluble, which delays the ability to build resistance to these drugs 19.
Many herbal extracts have been used in treating T. vaginalis [21,22,23] , and nanocomposites of some plants have also been used in the treatment[10,11]. From these plants C. colocynthis. The pulp of the fruits of these plants is used as sedatives and anti-inflammatories 24. It is recommended to use unripe fruits that do not contain seeds 25. “The pulp contains a bitter compound called colocynthene, a resin called colocynthin, colocynthia, pectin” 26. Of the active compounds that C. colocynthis, alkaloids, and this substance induces programmed death of parasite cells, changes in morphology, interruption of the cell cycle, and cytoplasmic vacuolization 27. Terpenes, an example of this is the (Capsaicin) terpene, have high biological activity in humans as it affects the nerves, and is a pain reliever, but it inhibits various types of pathogens, and its effectiveness against microorganisms is also attributed to its possession of a property. It is lipophilic and can link with the wall of a living cell and influence it through it inside it and work on forming complexes and connections with the components of the cell leading to the weakening of its vitality and destruction 28. Flavonoids are described as antibiotics, as studies have found that they have effectiveness against bacteria, fungi and viruses 29.
C. spinose also contains many active ingredients such as glycosides, myrosinase, rustic acid, caproic acid, pectic acid, saponin, alkaloid substances such as Stachydrineو And flavonoids such as Lerpenes and Flavonoids30, “Plants can produce a large amount of relatively small organic chemicals that are defined as secondary metabolites that lead to pharmacological or toxic effects”30. These compounds act as analgesics and anti-inflammatories, as well as inhibit the growth and reproduction of microorganisms 31. Also, the pulp contains more flavonoids and phenolic than the roots, which affect energy metabolism and plasma membrane permeability32, and alkaloids that can do the same action33.