|
|
| Year : 2014 | Volume
: 57
| Issue : 2 | Page : 356-357 |
|
| A modified fungal slide culture technique |
|
|
Arunava Kali, Sreenivasan Srirangaraj, M. V. Pravin Charles
Department of Microbiology, Mahatma Gandhi Medical College and Research Institute, Puducherry, India
Click here for correspondence address and email
| Date of Web Publication | 19-Jun-2014 |
|
|
 |
|
How to cite this article:
Kali A, Srirangaraj S, Pravin Charles MV. A modified fungal slide culture technique. Indian J Pathol Microbiol 2014;57:356-7 |
Sir,
Laboratory identification of pathogenic fungal agents essentially depends on demonstration of unique morphological characteristics. However, demonstration of intact fungal morphology is often not possible in widely practiced tease mount technique. We describe a simple modification of fungal slide culture method using cellophane tape, which combines the best of slide culture and cellophane tape preparation.
Slide culture set comprising of a glass petri plate and a slide was sterilized by hot air oven. A square agar block (7 mm × 7 mm × 3 mm) cut from sabouraud dextrose agar plate was placed on one-half of the slide, and a needlepoint inoculum of spores or mycelium was inoculated at the lateral surfaces of the agar block on all four sides. A drop of sterile distilled water was placed close to the agar. The agar block was snugly sealed to the slide with a 4 cm square strip of clean cellophane tape. Remaining half of the slide was wiped with methanol to decontaminate the surface from fungal spores. After its drying, same protocol was followed for inoculation of another fungal isolate on the other half of the slide [Figure 1]. The whole set was incubated at 25°C after closing the petri plate. When visible fungal growth on the cellophane tape was about 1-3 mm wide, the tape was lifted from the agar block, and lactophenol cotton blue (LPCB) mount was prepared on another slide for microscopic examination. | Figure 1: Two fungal isolates inoculated on two agar blocks placed on a microscopic slide and sealed with cellophane tape with a drop of sterile distilled water is close to the agar blocks
Click here to view |
Adhesive cellophane tape method is a fast and inexpensive technique, which obviates the need of teasing and provides better microscopic morphology. [1] In 1950, Riddell introduced a simple slide culture technique to obtain permanent stained intact mycological preparations. [2] Both these methods have their own advantages and limitations. Cellophane tape preparations are not suitable as permanent preparations due to quick drying and wrinkling of the tape. [3] On the other hand, owing to the open nature, slide culture ensures good aeration and moisture at the cost of contamination. It also requires conditions suitable for fungal growth viz. nutrition, aeration, moisture, and temperature.
We have tested 14 different fungal species from our stock cultures, which include fungi with slow growth rate viz. Trichophyton rubrum, Trichophyton schoenleinii, Trichophyton mentagrophyte, Epidermophyton floccosum and Microsporum gypseum as well as common rapid growing molds belonging to genus Mucor, Rhizopus, Syncephalastrum, Penicillium, Fusarium, and Aspergillus. We found this modified technique supported growth of both rapid and slow growing fungi without altered morphology, drying, and cross-contamination. There was no essential difference in growth characteristics in conventional and modified slide culture method. In fact, fungal structures were undistorted due to adhesive nature of cellophane tape. Moreover, cellophane tape LPCB mount had good microscopic clarity which is essential for fungal morphotyping. Due to self-containing nature of this sealed preparation, the risk of cross-contamination within the petri plate was diminished. More airspace within the sealed preparation was ensured by increasing thickness of agar block causing tenting of the cellophane membrane. Since this modification combines the best of slide culture and cellophane tape preparation, it could be beneficial in fungal identification.
 References | |  |
| 1. | Larone DH. Medically Important Fungi. 3 rd ed. Washington, D.C: ASM Press; 1995. 
|
| 2. | Riddell RW. Permanent stained mycological preparations obtained by slide culture. Mycologia 1950;42:265-70.
|
| 3. | Hughes AD, Lorusso GD, Greer DL. The 'double-layer tape prep': An improvement to a standard technique. J Med Microbiol 2004;53:455.
|
Correspondence Address:
Arunava Kali
Department of Microbiology, Mahatma Gandhi Medical College and Research Institute, Puducherry - 607 402
India
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/0377-4929.134756
[Figure 1] |
|
| This article has been cited by | | 1 |
Fungal-mediated deterioration and biodegradation study of low-density polyethylene (LDPE) isolated from municipal dump yard in Chennai, India |
|
| Merina Paul Das,Santosh Kumar,Jayabrata Das | | Energy, Ecology and Environment. 2018; 3(4): 229 | | [Pubmed] | [DOI] | | | 2 |
A modified micro chamber agar spot slide culture technique for microscopic examination of filamentous fungi |
|
| Peralam Yegneswaran Prakash,Kanika Bhargava | | Journal of Microbiological Methods. 2016; 123: 126 | | [Pubmed] | [DOI] | | | 3 |
Effects of gamma radiation on enzymatic production of lignolytic complex by filamentous fungi |
|
| Maria de Queiroz Baptista Nelania,Gomes Solidonio Evelyne,Virginia Ferreira de Arruda Flavia,Jonne Vieira de Melo Erik,Robson Neves Cavalcanti Filho Jose,Judith de Azevedo Callou Maira,de Cassia Mendonca de Miranda Rita,Colaco Waldeciro,Buarque de Gusmao Norma | | African Journal of Biotechnology. 2015; 14(7): 612 | | [Pubmed] | [DOI] | |
|
|
|
|
|
|
|
|
|
|
|
|
| Article Access Statistics | | | Viewed | 9623 | | | Printed | 97 | | | Emailed | 0 | | | PDF Downloaded | 201 | | | Comments | [Add] | | | Cited by others | 3 | | |
|
|
