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A novel, cheap and easy preparing selective medium for isolation of Pythium species
Sahar A Moharam1, Amna M Sadek1, Hani M A Abdelzaher2
1Department of Botany and Microbiology, Faculty of Science, Minia University, El-Minia, 61519, Egypt.
Abstract:
Isolation of Pythium and Globisporangium (Chromista, Oomycota) from their natural sources is very difficult due to the heavy contamination with other microbes, which hinders obtaining them in pure form. This study aims to develop a new, cheap and safe selective medium to isolate Pythium and Globisporangium species from their natural sources, despite the previous presence of more than 3 types of selective media including: VP3 (Vancomycin + PCNB + Penicillin + Pimaricin), NARM (Nystatin + Ampicillin + Rifampicin + Miconazole), and NARF (Nystatin + Ampicillin + Rifampicin + Fluazinam) used to isolate Oomycota. The elevated expense of antibiotics, along with alerts regarding potential contamination, toxicity, and tumor proliferation, renders them undesirable. Safe and inexpensive antibiotics from pharmaceutical sources were used with fixed formulations and concentrations. A selective medium was developed, called FANS (Fluconazole + Ampicillin + Nystatin + Sulbactam) with the following concentrations (1200 mg + 500 mg + 100,000 IU + 250 mg) respectively/L Corn Meal Agar promotes only the growth and formation of reproductive structures of Pythium and Globisporangium. On the other hand, these antibiotics completely inhibited the growth of other contaminated fungi and bacteria. Mycelial growth, zoospore formation, and oospore production for (Pythium aphanidermatum, Pythium oligandrum, and Globisporangium ultimum var. ultimum) were highest in FANS, followed by NARM, then NARF, while VP3 gave the lowest results compared to the control sample. It can be concluded that the use of FANS selective medium made the process of Pythium isolation easier, safer, and less expensive for worldwide researchers especially in developing and low-income countries.
Insights
A new, cost-effective selective medium, FANS (Fluconazole + Ampicillin + Nystatin + Sulbactam), simplifies isolating Pythium and Globisporangium. This method enhances Oomycota research by overcoming contamination issues and reducing expenses for global scientists.
Area of Science:
- Microbiology
- Mycology
- Plant Pathology
Background:
- Isolation of Pythium and Globisporangium (Oomycota) is challenging due to microbial contamination.
- Existing selective media (VP3, NARM, NARF) are expensive and pose safety concerns.
Purpose of the Study:
- To develop a novel, economical, and safe selective medium for isolating Pythium and Globisporangium species.
- To evaluate the efficacy of the new medium against existing methods.
Main Methods:
- A new selective medium, FANS (Fluconazole + Ampicillin + Nystatin + Sulbactam), was formulated using inexpensive pharmaceutical antibiotics.
- The FANS medium was tested for its ability to promote growth of Pythium and Globisporangium while inhibiting contaminants.
- Comparative analysis of FANS against VP3, NARM, and NARF was performed based on mycelial growth, zoospore, and oospore production.
Main Results:
- The FANS medium effectively isolated Pythium and Globisporangium, completely inhibiting bacterial and fungal contaminants.
- FANS demonstrated superior performance in promoting mycelial growth, zoospore, and oospore production compared to NARM, NARF, and VP3.
- The FANS medium proved to be a safe and cost-effective alternative for Oomycota isolation.
Conclusions:
- The FANS selective medium offers a simplified, safer, and more affordable method for Pythium and Globisporangium isolation.
- This development is particularly beneficial for researchers in developing and low-income countries.
- FANS enhances the accessibility and efficiency of Oomycota research globally.
