Abstract and References
Transactions on Science and Technology Vol. 5, No. 4, 233 - 238, 2018

A Laboratory Practicum on Screening for Lytic Bacteriophages from Soil Samples

Hasina Mohammed Mkwata, Armstrong Ighodalo Omoregie, Izzatie binti Musa, Jacynthia Suyuh, Phung Hui Yie, Ling Wen Sin, Lee Tung Tan, Peter Morin Nissom

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ABSTRACT
Bacteriophages are viruses that explicitly infect and lyse bacteria. They are ubiquitous and play vital roles in various biotechnological applications such as phage-based bacterial detection and bacteriophage therapy. This paper describes a simple bacteriophage screening protocol for teaching in secondary and tertiary institutions. This laboratory practicum was designed to enable students explore the presence of bacteriophages from soil samples. The tendency of many bacteriophages to produce plaques that are small or excessively turbid on agar plates limits their proper visualisation and enumeration. One percent of TTC (2,3,5-Triphenyltetrazolium chloride) was incorporated during the pour plate technique to enhance visibility of bacteriophage plaques on Petri dishes. The isolated bacteriophages were then amplified in growth medium containing targeted bacterial hosts. The results showed presence of six bacteriophages with distinctive plaque morphologies. Students were then taken through bacteriophage amplification and storage procedures. A survey was conducted to evaluate student’s learning experience both prior and after laboratory classes. Analysis from the survey showed that only 26% of students attempted all questions from the pre-laboratory bacteriophage practicum questionnaire whereas 74% failed the survey. On the other hand, 53% of the students attempted all questions for the post-laboratory phage practicum questionnaire and 16% failed the survey.

KEYWORDS: Bacteriophages; biocontrol; enrichment culture; phage isolation; plaque assay

REFERENCES

Allen, M. E. and Gyure, R. A. (2013). An undergraduate laboratory activity demonstrating bacteriophage specificity. Journal of Microbiology & Biology Education: JMBE, 14, 84.

Awang-Kanak, F., Masnoddin, M., Matawali, A., Daud, M. A., and Jumat, N. R. (2016). Difficulties experience by science foundation students on basic mendelian genetics topic: A preliminary study. Transactions on Science and Technology, 3(1-2), 283-290.

Casey, E., van Sinderen, D., and Mahony, J. (2018). In vitro characteristics of phages to guide ‘real life’phage therapy suitability. Viruses, 10(4), 163.

Chai, Y., Xiong, H., Ma, X., Cheng, L., Huang, G., and Zhang, Z. L. L. (2010). Molecular characterization, structural analysis and determination of host range of a novel bacteriophage lsb-1. Virology journal, 7, 255.

Fortier, L. C. and Moineau, S. (2009). Phage production and maintenance of stocks, including expected stock lifetimes. Methods in Molecular Biology, 501, 203-219.

Kumar, P. B., Kannan, M. M., and Quine, S. D. (2011). Litsea deccanensis ameliorates myocardial infarction in wistar rats: Evidence from biochemical histological studies. Journal of Young Pharmacists, 3, 287-296.

Lobocka, M., Hejnowicz, M. S., Gkagała, U., Weber-Dkabrowska, B., Wkegrzyn, G., and Dadlez, M. (2014). The first step to bacteriophage therapy – how to choose the correct phage, p. 378. In J. Borysowski, R. Mikedzybrodzki and A. Górski (eds.). Phage therapy: Current research and applications. Caister Academic Press, Norfolk, England.

Matsuzaki, S., Rashel, M., Uchiyama, J., Sakurai, S., Ujihara, T., Kuroda, M., Ikeuchi, M., Tani, T., Fujieda, M., and Wakiguchi, H. (2005). Bacteriophage therapy: A revitalized therapy against bacterial infectious diseases. Journal of infection and chemotherapy, 11, 211-219.

Masnoddin, M., Kiram, J. J., Matawali, A., and Jumat, N. R. (2018). Improving pre-university students’ understanding of basic plant tissue culture topic through laboratory teaching: a case study of UMS. Transactions on Science and Technology, 5(2), 53-57.

O'Sullivan, L., Buttimer, C., McAuliffe, O., Bolton, D., and Coffey, A. (2016). Bacteriophage-based tools: Recent advances and novel applications. F1000Research 2016, 5:2782

Sambrook, J. and Russell, D. W. (2001). Molecular cloning: A laboratory manual. Cold Spring Harbour Laboratory Press, New York, United States of America.