Mutagenicity of Makahiya (Mimosa Pudica) Decoction

Chapter I

BACKGROUND OF THIS STUDY

Plants have been a valuable source of natural basic products for maintaining individuals health, and the utilization of herbal vegetation as pharmacological treatments of diseases started out long ago. A lot of people who are below the poverty collection cannot afford the high cost of commercial fabricated drugs from drugstores and therefore choose to use local plants of their reach since these can be acquired easily, cheaply and entail easy prep and application (Racadio, 2008).

Mimosa pudica Linn, which is recognized as "Makahiya" in Filipino, is a popular ornamental seed, as its leaves fold up when activated by touch, temperature or wind flow (Balag-ey, 2009). It is considered as one common weed that abundantly increases within the Philippines. Aside from ornamental purposes, local individuals use the decoction of the flower for relief from common conditions, such as asthma, dysentery, dysmenorrhea, rheumatoid arthritis, coughing, fever, sore neck and far more (Castillo, et. al. 2005).

One of the major concerns nowadays on the basic safety of plant ingredients, is their mutagenicity. Mutagenicity identifies a substance or physical agent's capacity to cause mutations or genetic alterations. The recognition of plant components with chemicals or ingredients capable of inducing mutations is crucial since mutagenic ingredients can potentially induce malignancy (Ming, 2014). Makahiya has been uncovered to contain mimosine. Mimosine is a dangerous alkaloid that has an antimitotic activity that blocks the cell cycle and inhibits DNA synthesis (Xuan, et. al. , 2013) thus might cause genomic instability. This may be a factor that brings about the onset of genetic mutations.

So far, a report done by Jadhav et. al in 2013 shows that Mimosa pudica seeds, after S9 metabolic activation were found to be mutagenic and significant. Predicated on this premise, the research workers conducted an identical study, but rather than focusing on just the seed products of the herb, the research workers used the complete plant. Most of all, this study utilised the decoction remove of the vegetable since this is the one being used by the local people. Furthermore, the Muta-chromoplate set was used rather than the typical Ames test used by Jadhav et. al.

Objectives of the Study

This study directed to recognize the mutagenicity of Makahiya (Mimosa pudica) decoction and fresh ethanolic remove using Muta-ChromoplateTM system.

Specific Objectives

1. To determine the range of wells with color change per plate of Salmonella typhimurium TA100 strain using:

a. decoction with S-9 activation

b. decoction without S-9 activation

c. fresh ethanolic draw out with S-9 activation

d. fresh ethanolic extract without S-9 activation

2. To interpret the number of wells with color change per plate of Salmonella typhimurium TA100 strain utilizing 0. 05 level of significance in the next preparations:

a. decoction with S-9 activation

b. decoction without S-9 activation

c. fresh ethanolic draw out with S-9 activation

d. fresh ethanolic extract without S-9 activation

Null Hypothesis

1. No Hypothesis
2. No Hypothesis

Significance of the Study

This study is vital and beneficial to the following:

1. Society. Folks will be equipped with an increase of information and be aware of the mutagenicity of Makahiya as natural remedies. Most specifically, they'll be aware of the safe practices of the decoction prep of the said plant as it is often used in a decocted form.
2. Department of Health. With all the understanding of the analysis, the Division of Health will be able to utilize the studies and disseminate information to the people in the town of Davao regarding the mutagenicity of the utilization of Makahiya.
3. Researchers. The analysis may provide as an avenue in enhancing the skills of the researcher, specifically on methodical inquiry. In addition, this analysis may guide the research workers to explore further studies on the mutagenicity of Makahiya.
4. Future Research workers. This analysis may provide important knowledge, beliefs, and experience in performing clinical tests among future research workers if and when they plan to carry out a similar research. This analysis will be helpful when doing a medical inquiry on the issues or issues related to the mutagenicity of Makahiya.

Scope and Limitations

The scope of this experimental study was the determination of the mutagenicity of the decoction and fresh ethanolic extract of Makahiya. The natural herb was collected in Davao City and the ingredients were ready in Davao Medical University Foundation, Inc. lab, Dr. A. Gahol Avenue, Bajada, Davao City. There was no usage of animals as examining subjects. Alternatively, Muta-Chromo Plate Equipment with Salmonella tyhimirium TA100 pressure, which was based on the reverse-mutation test, known as "Ames Test" was used and provided by the EBPI Company. Also, the S9 enzyme activation was by using a crude liver remove provided in the system.

Theoretical Framework

The theoretical platform of this study was based on a test produced by Bruce Ames, commonly known as the Ames test. In the study conducted by Ames et al. in 1975 (as cited in Razak et al, 2007) they used a test that utilizes a bacterial strain of the Salmonella typhimurium as tool to discover mutations.

These S. typhimurium strains also have other characteristics that improve their ability to detect mutations. Mutation in the histidine biosynthesis gene makes the cell wall structure of the bacteria more permeable to large substances. Mutation in a gene accountable for proper excision and repair of DNA damage increases their awareness to mutagens.

The strains of S. typhimurium are known as auxotrophs which imply that they cannot produce a required nutrient. The test organism cannot synthesize the amino acid histidine, thus, you won't expand unless the nutrient comes in the growth press. Auxotrophs are usually produced as a result of the mutation occurring in a prototroph, a bacterium that is able to synthesize the particular nutrient.

The Ames' test decides the ability of any tested compound to cause a reversal, also known as a back-mutation of these auxotrophs to the original prototrophic state. During the test, auxotrophs are cultivated in glucose-minimal salts agar plates that contained all required nutrition but only trace levels of histidine and biotin. The auxotrophs are able to grow for a number of generations before histidine in the media was exhausted. This time they will stop growing unless they have suffered a back-mutation that has restored their ability to synthesize histidine.

Suspected mutagenic chemicals were tested for his or her ability to induce back again- mutations by positioning set on the surface of the minimal agar plates previously inoculated with the auxotroph. The test substance diffused in to the surrounding mass media and, if mutagenic, would cause back-mutations which would then allow growth into obvious colonies. The more revertants observed near the test substance, relative to experimental controls, a lot more mutagenic the chemical was apt to be.

Conceptual Framework

The study targeted to determine the mutagenic activity of decoction and fresh ethanolic draw out of Makahiya on Salmonella typhimurium TA100 tension which involved substitution, addition, or deletion of one or a few DNA base pairs. The unbiased variables were the decoction and fresh ethanolic extract of Makahiya. The dependent variable was the amount of wells with color change per bowl of Salmonella typhimurium TA100 strain, with and without the addition of the S9 activation enzyme.

Figure 1. 1 Conceptual Construction of the study

Definition of Terms

Ames test. A test used to look for the mutagenic potential of any substance based on the mutation rate of bacterias that were subjected to the compound.

Decoction. A method that involves boiling to be able to extract a plant material.

Fresh ethanolic remove. Extract collected from M. pudica by using dilution with 95% ethyl alcohol followed by rotary evaporation process.

Makahiya. Scientific name, Mimosa pudica. The plant sample used in this review.

Muta-ChromePlateTM. A 96-well micro-plate version of the Salmonella typhimurium "Ames Test

Mutagenicity. The capacity to generate mutation.

Salmonella typhimurium. The pressure used for the Ames test. It carries mutant gene that prevents them from synthesizing the fundamental amino acid histidine from the materials in standard bacterial culture medium

S-9 enzyme. S-9 is a crude liver enzyme extract useful to mimic mammalian metabolism. This enzyme is bought together with the kit.