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The counterstaining potential of Bixa orellana (Atchuete) seed extract for Escherichia coli and Staphylococcus aureus.


Charmaine P. Avila,
Miguel Elijah T. Bibera,
Angelie Monique L. Gurrea,
Carl A. Luzano,
Christine Megan D. Sesaldo,
Quennie Liz T. Tumanda,
Samantha Faith Y. Tan,
Brian  M. Denney

Related Institution

College of Allied Medical Sciences - Medical Technology Department - Cebu Doctors' University

Publication Information

Publication Type
Thesis Degree
Medical Technology
Publication Date
October 2015


        Staining of bacterial organisms is the first and most essential procedure to begin the myriad of diagnostic  methodologies used in identifying the causative organism for all diseases of bacterial origin. Medical technologists and laboratorians do not haphazardly perform all the available tests used in the identification of bacterial pathogens for this would be a waste of time, monetary resources and possibly even the lives of patients. The study aimed to produce a natural dye that is simple and readily made, safe to handle and dispose of in the laboratory setting. Thus, the researchers aimed to discover if Bixa orellana (Atchuete) seed extract can serve as an alternative counterstain to Safranin, which can be made readily available, is less hazardous to health and safety, efficient in staining bacteria, and at the same time may improve the livelihood and income generation of local farmers and agriculturists alike. Although both from organic sources, the preparation of commercial Safranin is truly different and may result to various potential hazards in the context of potential carcinogenic effects of vapors and the potential to behave as a volatile oxidizer. The researchers smeared eightyeight (88) slides with the chosen representative organisms, forty-four (44) of which were smeared with Escherichia coli and the other forty-four (44) were smeared with Staphylococcus aureus. The researchers duplicated this set-up to obtain two (2) groups, one group for the control and the other group subject to treatment. Slides of the control group were subjected to the traditional method of gram staining while the experimental group was treated similarly using the same gram staining process but with the exception of the counterstain used, Safranin, for this was substituted with Bixa orellana (Atchuete). Three (3) highly experienced registered medical technologists were respectfully tasked to evaluate the one hundred and seventy six (176) slides for adequacy of staining.

        The raw data gathered was collated and subjected to statistical treatment and results are as follows: one hundred percent (100%) of the slides designated as control were evaluated as having adequate and positive staining for both Escherichia coli and Staphylococcus aureus. None of the smeared slides of the Escherichia coli experimental group exhibited an adequate staining result, thus deemed negative, while, 9.1% of the Staphylococcus aureus experimental group slides gave pleasing results. Although the latter gave promising results, the number of the positively stained Staphylococcus aureus smears are not at all statistically significant in the context of this study.

        Results of the experimentation conclude that Bixa orellana does not have counterstaining potential in the Gram staining process of Escherichia coli and Staphylococcus aureus. Based on this conclusion, the researchers would like to recommend the use of accentuators, mordants, and other additive substances to enhance the staining potential of the Bixa orellana stain, the participation of other representative organisms such as fungi, parasites and other species of bacteria, the aging of the seed extract naturally or by oxidizers and the performance of different dilutions to adjust the concentration of the staining solution.


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