To culture bacteria or fungi, you need the following materials. But, it takes time, some fungi take months to sporulate. The ability of fungi to invade plant and animal tissue was observed in early 19th century but the first documented animal infection by any fungus was made by bassi, who in 1835 studied the muscardine disease of silkworm and. The process of transferring hyphae to a slide breaks up the structure of hyphae and sporangiophores so that identification becomes very difficult. Modifiedfaster and easier identification of dermatophytes cotton blue stain. View lab report exercise 19 fungal culture from bio 120 at university of the philippines visayas. Preparation of permanent stained slide cultures jama. It is a costeffective, rapid method for the identification of fungal isolates based on their morphological and microscopic characteristics. Pdf evaluation of an alternative slide culture technique. An easy, cheap, and rapid agar plate assisted slide culture technique is developed which facilitates the study of invitro mycoparasitism with ease. Slide culture technique was use for proper identification of the filamentous fungiand incubation was at least four 4 six 6 days.
Slide culture technique preparation of a slide culture can be useful in the study and identification of an unknown fungal isolate. Place a slide, concave side up, with a coverslip to one side of the concavity, on the glass rod in the petri dish. Priority is given to contributions likely to be of interest to a wide international audience. While the sticky tape preparation is quick and easy to perform, it can be disruptive, with salient features altered or destroyed. A modified slide culture method which combines advantages found in several slide culture methods is described. Safe, lowdistortion tape touch method for fungal slide mounts.
Adaptation of a polystyrene plate slide is extremely well suited for the simultaneous macroscopic and microscopic diagnosis of fungi. To become familiar with mycological culture techniques. A doubleoverglass technique used for preparing permanent slides of marine fungal spores is herein adapted to the standard slide culture technique. Cultivationculturing of bacteria a microbial culture, is a method of multiplying microorganisms by letting them reproduce in predetermined culture media under controlled labor. With a sterile pasteur pipette, add one or two drops of cooled sabouraud agar ca. The present paper describes an improved slide culture technique which is relatively easy to oarry out and is useful for practical diagnostic work in mycology, as well as for classroom teaching.
Document bemire ed 174 459 title mycology guidebook. To have more practice in serial dilutions and calculations. Readytouse, diagnostic slide culture system for identifying fungi the mycovue system provides the laboratorian with a standardized, comprehensive method that eliminates timeconsuming preparations and technical difficulties encountered with the classical slide culture technique. A modified technique developed by harris in 1986 utilized a layer of water agar to provide humidity. Dec 16, 2018 hey friends im medical laboratory scientist. The use of the slide culture technique is well known among dermatologists as a valuable diagnostic and teaching aid in mycology. Slide culture technique this technique is the best technique for the microscopic examination of mold cultures. Two general types of culture media are essential to ensure the primary recovery of all clinically significant fungi from clinical specimens. A coverslip culture technique for the growth of fungi, with subsequent preparation of permanent mounts for microscopic examination, is described. A simple modification of this method using a single agar plate is described below. Long viability ensured protectionagainst contamination works well for allfungidisadvantages expensive technique complex technique many copies areneeded as someisolates fail to survive viability needs manytrials. However it takes time some fungi take months to sporulate, and even after months, only sterile hyphae can be seen. An improved slide culture technique for the study and.
To visualize and identify the structural components of fungi. This simplified technique allows examination at different stages of development of the fungus, or can be used in the preparation of large numbers of similar mounts for teaching purposes. Media will affect colony morphology and color, whether. A new agar plate assisted slide culture technique to study. This technique allows the intact morphology of the fungus to be seen under the microscope. I am facing a problem in identification of fungus under microscope. It simplifies the slide culture method by providing all the. Jackson and colleagues 1991 used this methodology to screen for potential biocontrol capacities of numerous soil isolates against sclerotium cepivorum. A technique for the slide culture of fungi science. May 27, 2011 slide culture method is a rapid method of preparing fungal colonies for examination and identification.
Fungal slide cultures made by this new method will last indefinitely. Precise methods of identification of moulds accurate identification of. Evaluation of an alternative slide culture technique for the morphological identification of fungal species article pdf available august 2012 with 1,956 reads how we measure reads. Fungi are identified mostly by close examination of its morphology and the characteristics it possess. Overview of fungal identification methods and strategies. Culture media for fungi introduction a wide range of media are used for growing fungi. The simple method of slide culturing used widely is described here, which permits fungi to be studied virtually insitu with as little disturbance as possible.
It is not a rapid technique, but it is unsurpassed as a routine means for studying the. Basic slide culture as a morphological identification was known as the most common method for the identification of pathogenic mold fungi. Pravin charles department of microbiology, mahatma gandhi medical college and research institute, puducherry, india click here for correspondence address and email. In slide cultures, we are growing the fungi directly on the slide on a thin film of agar. An improved technique for making permanent slide cultures. None of these methods is suitable for rapid identification of pathogenic fungi. Pdf morphological identification is still the most commonly used method for the identification of pathogenic fungi, with slide culture being the most.
Evaluation of an alternative slide culture technique for the. After adequate growth has occurred, the slide culture is disassembled. Lactophenol blue was use to stain the fungi growth on the slide and the cover slip which was viewed under the light microscope. Slide culture technique for fungi as soon as most ofthe alcohol has evaporated, take the cover slip from the glass slide using a forceps and place it with the growth side down, on the drop of lactophenol cotton blue stain on the slide. Johnson a department of bacteriology and immunology, baylor university college of medicine, houston, texas. Inoculating loop for bacteria, yeasts, and fungi with abundant spores. After adequate growth has occurred, the slide culture is disassembled and mounted in a conventional manner. Objectives to demonstrate good aseptic technique in culture transfer or inoculation and in handling sterile materials. With a toothpick, add petroleum jelly to three sides surrounding the concavity of each slide. Sterile slides or cover glasses are placed on a layer of filter paper in a petri dish.
For some purposes, moreelaborate specimenchambersarenecessary. When the alcohol evaporates, place the cover glass on the slide having the lactophenol cotton blue. Lutchmeah and cooke 1984 adopted a slide culture technique to observe the parasitic action of pythium fungicolous fungi oligandrum microscopically. Introduction to mycology the term mycology is derived from greek word mykes meaning mushroom. What is the best method for the identification of fungus. Common fungal culture media and their uses microbeonline. An improved technique for making permanent slide cultures of. Plateslide for the culture and morphological observation fungi. What is the main advantage of using the slide culture technique for identifying molds. Slide culture in mycology freelance clinical microbiologist. One medium should be nonselective such as brain heart infusion agar.
Several tape flags may be prepared at once if multiple tape touch mounts are to be made. Assist with the recognition of fungi recovered on culture media that are strictly pathogenic fungi. Techniques for the microscopic examination of fungal culture. Morphological identification is still the most commonly used method for the identification of pathogenic fungi, with slide culture being the most widely used. The time and the nature of the block of agar are determinant factors. A block of inoculated nutrient agar sandwiched between two sterile cover glasses is placed in a plastic petri dish containing water agar. Easier setup than slide culture method position of reproductive structures to hyphae are maintained disadvantagecoverslip easily breaks cottony fungi grow past edges waiting period for incubation on potato dextrose agar platefungal elements are vegetative. This video has information about slide culture technique for fungi. Disinfectant solution such as 70% ethanol, 4% household bleach solution, or lysol. An improved slide culture technique for the study and identification of pathogenic fungi 1 edwin a. A drop of melted culture medium is placed on the slide or cover slip and inoculated.
Place a coverslip over the concave portion of each slide so that it is completely sealed. For these reasons, we developed a modification of the adhesive tape technique whichfacilitates and improves the microscopic examination ofmoldcultures. The first page of the pdf of this article appears above. A coverslip culture technique for preparing permanent fungus. In the past, most fungi cause skin infections or cosmetic infections, where bacteria and viruses cause serious fatal diseases, so there was no interest of studying fungi. With the help of this technique mycoparasitism by a strain of trichoderma. Lutchmeah and cooke 1984 adopted a slideculture technique to observe the parasitic action of pythium.
There is amyriad of infections causedby these agents, rangingfrom superficial mycoses todiseases with highmorbidity and mortalityrates. Nov 06, 2016 cultivation of bacteria and culture methods 1. I use lactophenol cotton blue flooding over slide where fungus is placed from fungal plate. Morphology helps inbasic identificationmany pathogenic fungi maybe recognized. For studies of geotropism in fungi, the modified slide culture chamber may be conveniently placed on edge and. We describe a modified and easy to perform micro chamber agar spot slide culture technique. The procedure involves the use of a device pelikan rollfix. Nov 04, 2011 slide culture technique preparation of a slide culture can be useful in the study and identification of an unknown fungal isolate.
Although the most common technique used in mycology, identification is often difficult by tease mount method because of the dislodgement of. Importance of fungal infectionsfungi have a greatimportance in the field ofmicrobiology, for they affectall living beings. From the slide culture remove the cover glass, and discard the agar block. In 1980, when hiv infection was discovered, increasing number of immunocompromizing conditions, they found that fungi produce fatal diseases. Pdf modified slide culture method for faster and easier. Most mycologists develop preferences for certain types of media based on experience and peculiarities of the type of fungi that are routinely grown.
With forceps, stand the slide upright inside petri dish until the agar solidifies, as illustrated below. Slide culture method is a rapid method of preparing fungal colonies for examination and identification. Slide culture technique definition of slide culture. One of the disadvantages of this procedure has been the inability to maintain permanent stained mounts for future reference in a teaching program or when attempting to identify an unknown fungus. For accurate identification of fungi, it is required that the precise arrangement of the conidiophores and the way in which the spores are produced is essential. The simple method of slide culturing used widely is described here, which permits fungi to be studied virtually. Slide culture technique and lactophenol cotton blue staining. Now add 95% ethanol to the hyphae on the cover glass. Pdf an improved slide culture technique for the study.
Riddels simple method of slide culturing mycologia 42. The slide culture technique aids in the study of undisturbed microscopic morphological details of filamentous fungi. Name three stains or reagents that may be used to facilitate the microscopic detection of fungi in clinical samples. Techniques for bacteria and fungi fisher scientific. The contents can then be transferred to a drop of sterilized water on a small glass container or slide in order to provide a spore suspension. Simplified technique for growing fungi in slide culture all current methods for growing fungi in slide cultures are based on the principle of incubation in a moist chamber. This method preserved the morphological features relatively undisturbed compared with adhesive tape preparations. With a bit of skill, you can also use sticky tape to sample a fungus growing in a test tube. Pdf basic slide culture as a morphological identification was known as the most common method for the identification of pathogenic mold fungi. The colekendrick culture chamber is still one ofthe best chambersfor timelapse studies ofconidiogenesis 2. Slide culture technique and lactophenol cotton blue. To be able to differentiate between different enumeration techniques and learn when each should be used. Isolation and identification of fungi from packaged and. What is the main advantage of using the slide culture.
As fungi grow directly on the slide on a thin film of agar, there is no need to remove a portion of the fungus from a culture plate and transfer it to the slide. Objectives to learn the different techniques used to count the number of microorganisms in a sample. Slide culture technique for accurate identification of fungi, it is required that the precise arrangement of the conidiophores and the way in which the spores are produced is essential. This method permits the fungi to be studied virtually insitu with as little disturbance as. Slide culture of fungithis procedure, whichallows fungal growthto be examinedundisturbed.
Pdf an improved slide culture technique for the study and. By doing this, there is no need to remove a portion of the fungus from a culture plate and transfer it to the slide. Semiselective media for general use selective media for specific. To demonstrate skill in isolation of organisms from a mixed culture using selective and differential media. Pdf evaluation of an alternative slide culture technique for the. Fungal biology is the international research journal of the british mycological society. The isolation, culture, and microscopic examination of molds require the use of suitable selective media and special microscopic slide techniques. However it takes time some fungi take months to sporulate, and. Slide culture is the best method for preserving the actual structure of a fungus.
This technique obviates the disorientation of identifying structures whencultures are mechanically disaggregated during preparation ofslides for microscopy. Efficacy of iodineglycerol versus lactophenol cotton blue. A modified micro chamber agar spot slide culture technique. Simplified technique for growing fungi in slide culture. Fungi with cupshaped fruiting bodies ascomycetes with apothecia. Slide culture technique for fungi amrita university youtube. To prepare a mount, hold a tape flag by the applicator stick handle and touch the tape to a fungal colony in a culture dish opened in a biological safety cabinet.
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