Fungal Biotechnology

Fungal Biotechnology

  • Christine Schimek added an answer:
    How long are fungal spore cultures stable in physiological salt solution stored in a fridge (+4C)?

    How long are fungal spore cultures stable in physiological salt solution stored in a fridge (+4C)?

    Christine Schimek · SRH FernHochschule

    I have also had some experience with another technique for mitospores of mucoralean zygomycetes: a droplet of a concentrated spore suspension put onto a small piece of sterile filter paper, allowed to dry, and then wrapped tightly in aluminium foil - but they might also be stored in a small vessel. Some spores stored in this way germinated even after up to two or three years storage in a refrigerator.

  • Muinat Olanike Kazeem added an answer:
    How can we calculate CFU/ g- 1 (Colony forming unit) of tissue burden candidasis for experimental study in mice?

    This is method use for this study:
    Lungs, brains, spleens, livers and kidneys were aseptically removed and approximately half of each organ was weighed and homogenized in 1 ml of sterile saline. Serial 10-fold dilutions of the homogenates were inoculated on SDA, incubated at 35 C and examined daily for 3 days. The number of CFU/g-1 of tissue was calculated for each sample.
    May I help me to answer these questions?
    1. Is it necessary we weighed organ?
    2. What is exactly meaning of Serial 10-fold dilutions?
    3. How can we calculate CFU? Any protocol or methods for this?

    Muinat Olanike Kazeem · University of Ilorin

    All your comments were absolutely right. Thanks

  • Nirmal Sahay added an answer:
    How can I perform pathogenicity test for fungi using petri plate method ?

    i am working on pathogenic diseases of some medicinal plants and need to carry out pathogenicity test using petri plate method.

    Nirmal Sahay · Society for Research and Initiatives for Sustainable Technologies and Institutions

    The simplest one is to 1st prove the Kochs Postulate. I need not to explain it you must be aware of it. If it is positive, then identify the fungus first. After its identification you will come to know through literature survey how important your identified pathogen is and what is its potential in disease severity. 

    There could be many parameters to know the potential of Pathogenicity

    a. Physical,

    b. Chemical and

    c. Biochemical

    but 1st prove whether it is pathogenic or non pathogenic in invivo or invitro [in Petriplate by germinating the seeds or through vegetative propagation (tissue culture) ] condition.

  • Michael Seegers added an answer:
    Where can I get spores of Cordyceps sinensis and C. militaris?

    Can anyone suggest me how to buy the fresh spores of Cordyceps sinensis and C. militaris fungi?Or please let me know any lab which is working with these fungi?

    I am very interested in these species, since they are high value for medicine.

    Thank you in advanced.

    Dr. John Holliday is the world's largest producer with the best quality including DNA verification. See http://www.alohamedicinals.com/

  • Marc Collins asked a question:
    What does the growth morphology of a fungus in a shaken liquid medium say about the medium?

    I want to achieve maximum cell biomass over 72 h in a shaken flask inoculated with a fungus. Some grow really well but others don't particularly Curvularia lunata and Rhizopus Stolonifer. Are there any studies dealing primarily with fungal growth in shaken Liquid media?

  • Shrvan Kumar added an answer:
    Could anyone suggest why I found no bands in my gel although I extracted DNA with high purity (as shown by nanodrop)?

    I did extract by CTAB and I tried phenol. DNA from fungi.

    Shrvan Kumar · IAS, BHU-VARANASI (UP)-221005

    dear sir firstly,

    1. Check pH meter and calibrate it.
    2. Check pH extraction buffer.
    3. isolated DNA in pure form diluted in RNase free water.
    4. Check your Nanodrop and calibrate it.
    5. 1 %  agarose solution and check pH.
    6. Gel run at 60 V for 30 minutes with dye.
    7. Check own Geldoc system and adjust the UV light
  • Adnan Uzunovic added an answer:
    How do I homogenise fungal mycelium from plates or liquid cultures to make an inoculum?

    Often in literature it is only mentioned that fungal cultures are homogenized but rarely it mentions the  time, type of homogenizer etc. There surely must be optimal choices as some bad choices as different homogenizers have different rpm power, blades can be of different quality and can be blunt of sharp. How long to homogenize to get best  propagules (mucelial fragments, blastospores, other spores..)  and the least of dead shattered mycelial fragments. I am working with basidiomycetes (wood decay fungi) . Any suggestions and personal experimences?

    Adnan Uzunovic · FPI Innovations

    Thanks Alessandra. Doing a pilot study on homogenisation for one's particular fungus certainly makes sense to avoid guessing game  and I am already starting a small pilot study.

  • Ramona Pezzotta added an answer:
    Is the E-test on SAB agar for the Scedosporium apiospermum reliable?

    I had to perform a MIC test on Scedosporium apiospermum last week (isolated from a BAL). Unfortunatly I couldn't use the specific and required media I was suppose to (potato's agar to select the conidia and R.P.M.I. for the E-test). I was in a sort of rush and then I decided to perform the E-test on Sabourad agar. I know that it's not correct, but I need to know if the results I obtained are reliable or not. Can I use them as an indication for the therapy?

    I tested Voriconazole (which looks Sensible) Itraconazole (which has a strange MIC of 0.5 but I think is dose-related) Ketoconazole (sensible as VOR) and Caspofungin (that's surely resistant).

    Any suggestion? Can I confirm to the clinical doctor this results?

    (He already started a VOR therapy, and the patient seems to feel, after few days, better. But the doctor wants a confirm anyway).

    Ramona Pezzotta · Spedali Civili di Brescia

    I definitely will .

    Thank you.

  • Woo-Sik Jo added an answer:
    Has anyone used herbarium material for antioxidant extraction of fungal fruitbodies?

    I would like to know how old can the fungal fruiting body be for detection and screening of antioxidants. Has anyone tried this with herbarium material? Does the preservation method affect the contents?

    Woo-Sik Jo · S. KOREA / www.gba.go.kr

    Dear. Dr. Masoomeh Ghobad-Nejhad 

    Please refer to the attached document research.

    Sincerely Yours   Woo-Sik Jo

    2015.05.12.TUE. 00:25[KOR]

  • Heba Shawky Mohamed added an answer:
    What is the best way of starting to look for commercially important metabolites in a newly characterized fungus?

    1. We can not order standards for all important metabolites.

    2. What type of Bioassays should be conducted to know if the fungi are producing any important metabolites?

    3. From Chemical or molecular approach, which one should be selected first? As both have their advantages and disadvantages. 

    Heba Shawky Mohamed · Egyptian Atomic Energy Authority

    I suggest to start with the chemical characterization firstly to know the major metabolites produced by this fungus. This is carried out by cultivating the fungus in a liguid rich medium as PDB or YSB media,then extract using different solvent systems to obtain the crude extract using rotary evaporator. this extravt can be then characterized through different techniques of chromatography such as column chromatography or GC/MS for recognition of these compounds. this can help you for chracterization.

  • Miss Biology added an answer:
    How can I read the OD from minimal salt solution plus with oil and fungus, for biodegradation of petroleum by using studies of isolated fungi?

    The 10ml minimal salt solution + 2ml of petrol(oil) and selected fungi was put in the test tube and being incubated in the incubator shaker(120rpm) for 20 days with 28 degree celsius, and the reading of Optical Density will be taken in 5 days interval at wavelength 540nm..in order to know the degradation rate of oil using fungi. Is this the best way to study the biodegradation rate of oil using isolated fungi?

    Miss Biology · Universiti Teknologi MARA

    Thank  you very much Eleni Xenofontos for answering my questions with the useful suggestions. I really appreciate it.

  • Dyoni Matias de Oliveira added an answer:
    Can fungi utilize the degradative products of lignin as their carbon sources?

    Or it degrades lignin to reach cellulose or hemicellulose inside?

    Dyoni Matias de Oliveira · Universidade Estadual de Maringá

    Yes, lignin degradation products can be used as a carbon source for ligninolytic fungi.

  • Partha Pratim Dhar added an answer:
    Is it wrong if I extracted mycelium fungi using ethyl acetate?

    I am doing my research about secondary metabolite from Sponge Associated Fungi. I have cultivated my fungi in broth medium, then I separated the mycelium from the media. Then, I extracted the mycelium using Ethy Acetat after disrupted it. Someone said for me, it is uncommon for the extraction.
    Thank you.

    Partha Pratim Dhar · King Saud University

    I am agreed to Ravi kant.

  • Kuldeep Patel added an answer:
    What techniques are available to study the viability of arbuscular mycorrhizal spores?

    I am using Tetrazolium (INT) as a viability stain for AMF spores but I am finding some inconsistencies in the results I obtain. Any other technique that I could use to contrast my results with INT? Thanks for your answers.

    Kuldeep Patel · BACL

    We are using MTT staining of mycorrhiza spores and IP by root staining

  • Andrej Gregori added an answer:
    Is anyone willing to share Monascus purpureus cultures?

    We are conducting cultivation of Monascus purpureus on solid media in order to reduce citrinin and increase monakolin production. We are having quite some problems finding M. purpureus cultures. Is anyone willing to share/exchange? We have many basidiomicetes (especially lignivorous) - Ganoderma, Lentinula, Pleurotus, Grifola, Trametes etc. in our collection which we can offer to you.

    Andrej Gregori · Zavod za Naravoslovje

    Dear Ravi,

    Thank you for your reply. Is TISTR 3541 the strain you are willing to share?

  • Hani Antoun added an answer:
    Can I use PDA medium with ampicillin to isolate endophytic fungi from macerated roots?

    Sterile-surface roots were macerated. From this material I isolated dozens of endophytic bacteria. Now I want to take this material to isolate endophytic fungi. I plan to use PDA medium with the only antibiotic that I have, ampicillin. Would it be okay supplement the PDA with ampicillin for this purpose?

  • Afreen Haider added an answer:
    How can I get the ribosomes in a total nucleic acid extraction?

    I need to do total nucleic extraction from fungi out of clinical samples. At the moment we are using bead beating followed by a total nucleic extraction protocol on the Roche MagNA pure compact. Quantification of spores going into the extraction (can't really quantify mycelia but have tried those too) and subsequent PCR (with a ribosomal target) suggests we are getting a good yield of DNA from this procedure. However, when we do reverse transcription PCR to pick up the RNA in the ribosomes as a target too. We are not seeing much more of a PCR product. Is there a way to ensure ribosomal RNA gets extracted in total nucleic extraction protocol.?

    Afreen Haider · Central Drug Research Institute

    You should first check the integrity of your RNA i.e. on agarose gel you should obtain intact bands for rRNA (3 prominent bands where the top two should be more intense than the lower one). If you see smears in the gel then your RNA is probably degradng hence you are not getting RT-PCR. But If that is fine and you are also using the same primer and buffers for both your normal DNA PCR and RT-PCR, then the only thing is that the RT step is not working. Are you incorporating a 65 degrees melting step before you incubate with RT? this removes secondary structures that can inhibit your RT reaction.

  • Ulrike Martens added an answer:
    Do you know any recipe for cellulose agar media?

    I made up cellulose agar media to grow fungi but the only growth was bacteria, some gram negs. I thought the salts would inhibit the bacteria and select for fungi that broke down cellulose

    Cellulose  (cmc) 5.0g

    NaNO3 1.0g

    K2HPO4 1.8g

    MgSO4 7H2O 0.9g

    KCl 0.5g

    Agar 20g

    dH2O 1000ml

    Any suggestions on tweaks to be made?

    Ulrike Martens · University of Greifswald

    I also used the described method. To test the cellulolytic activity add CMC in a concentration of 0,1 %. After incubation period cover the plates with aqueous solution of Congo Red (1 mg/ml) for 15 minutes. Then float it off with a solution of NaCl (1 M). Cleared zones indicate a cellulolytic activity.

  • Emmanuel Manirafasha added an answer:
    Anyone know where I can conduct the supercritical fluid extraction in Saskatoon or in Saskatchewan?

    I am trying to extract Fusarium mycotoxins from some grain samples using supercritical fluid extraction. Does anyone know any institution/facility have the capacity or experience for this process? I am a graduate student in University of Saskatchewan. If the facility close to Saskatoon or within Saskatchewan will be prefect. Any place in Canada will be great

    Emmanuel Manirafasha · Xiamen University

    I know one Institution in China, College of Chemistry and Chemical Engineering, Department of Chemical and Biochemical Engineering, Xiamen University 

  • Can Hemileia vastatrix grow on agar?

    I want to know if Hamileia vastatrix can grow in the lab on artificial media. And if yes what type of agar?

    Einar Martinez de la Parte · Instituto de Investigaciones de Sanidad Vegeta

    I agree with D. Andrivon, most of the people that works with H. vastatrix maintain the pathogen on detached coffeee leaves. Despite the article of Bakala et al. (1975), recent works with this rust do not used culture media for maintain H. vastatrix inoculum in the lab.

    best regards

    Einar

  • Shivakumar Banakar added an answer:
    How can I purify and characterize the extracted natural pigments from the fungi?

    I am a Postdoctoral Fellow and having a few difficulties in purifying the natural fungal pigments.

    I extracted pigments (colors) in liquid medium (Smf),  and came to know that they are soluble in water. I tried a few solvents, but they are insoluble.

    I want to know the purity of the natural compound, either one compound or multiple compounds and I want to characterise all.

    Please help me regarding this.

    Shivakumar Banakar · North West University South Africa

    Thank you for your reply Prashanta Kumar pal.

    Can you mention any of method to try for the suitability of the solvents.

    Initial samples were sent for the PREP HPLC, but they were told that tailing was observed after the running, because I do not have the facility so.

  • Hideo Toyama added an answer:
    Do you know The Mega-mushroom?

    Are there someone who study about the Mega-mushroom?

    I have a interest in the lignin and cellulose degrading ability of this fungus.

    I hope to exchange informations about this fungus.

    Biofuel is quite easy for this fungus???

    45lb Mushroom found in Mexico

    http://www.supersizedmeals.com/food/article.php/20070715-Giant_Mexican_Mushroom/print

    Hideo Toyama · Minamikyushu University

    Dear Ricardo Valenzuela

    Thank you very much for your informations.

    I begin to study about this monster.

    Thank you very mucj again!

    Sincerely

    Hideo Toyama, Ph.D.

  • Phurpa Wangchuk added an answer:
    Is anyone working with the isolation and characterisation of molecules from worms and insect fungi?

    The sources of drug discoveries has been a synthetic chemical pools and the natural resources including microbes, proteins, plants and animals. The worms especially the intestinal helminths has been neglected and I am wondering if anyone has any experience in this area including isolation, characterisation and biological activies-protocols and findings would be appreciated?

    Phurpa Wangchuk · James Cook University/University of Wollongong, Australia

    Thank you all for your comments. Prof Russell, how have you been doing? I saw one of your paper on insect fungi. I have made a request and would appreciate if you could also give a link to related works of yours.  Thank you.

  • Juraj Medo added an answer:
    How can I design a primer with GC-clamp attached to ITS ?

    I have read the articel that is suggested by Kuldeep Patel.Thank you so much.

    I carried out DGGE to study microflora on 18S RNA using primer (NS1 and GC fung) based on Technical report from National Institute for Agro-Environmental Science japan (NIAES).

    Currently,I would like to identify spesies from DNA band of DGGE

    I would like to use ITS1 and ITS4 for DGGE to study microflora of fungi, I would like to ask how to attach GC clamp on the primer, and how long  the GC sequence?

    Anybody could help me please?

    Thank you

    Beware on double bands, It is common if Muyzer`s GC clamp is attached to ITS primers. In my opinion IGS primers gave nicer gels

  • Hani Antoun added an answer:
    Is staining for mycorrhizae better with blue-black ink?

    I have been staining Celastrus orbiculatus roots and I have noticed that there are a variety of things that are being stained besides the mycorrhizae (hyphae, vesicles and arbuscules). They could either be dark septate endophtyes, microesclerotia, or the cell wall of the plant cells in the cortex being stained. I figured changing colors could help decipher the difference, but am reluctant to switch to blue ink because I have seen images of unclear samples of arbuscules when using blue ink. Does anyone have a particular preference or insight when using blue-black ink?

  • Mukesh Dubey added an answer:
    Why does Agrobacterium tumefaciens develop visible clumps (agglutination) in liquid LB medium?
    What is the solution to stop this? And is it okay to use clumped cells for transformation?
    Mukesh Dubey · Swedish University of Agricultural Sciences

    Hi manoj !

    I agree with you !

  • Gh V Chowdary added an answer:
    What is a suitable method for mass production of amylase using fungi?

    First I would like to start at laboratory level.

    Gh V Chowdary · horizon college of pharmacy

    For comnercial production it is advised to use rice husk/wheat bran by koji room method. Plenty of methods available pl go through lit. But do not use synthetic media due to lack of economical viability.

  • Ankur K. Anand added an answer:
    Is there any traditional microbial method for speed up growth of microorganism (MLT) in Industrial basis?

    Is there any traditional microbial method for speed up growth of microorganism (MLT) in Industrial basis? For Saving Days of released the result.

    As traditinaly we are doing incubation for fungal 5-7 days. What are the alternative we can use?

    Ankur K. Anand · Avon Products, Inc

    Thanks Christoph & Bhukya,

    As I am working on C. albicans & A. brasilliensis.

  • Is fungal crude extract suitable for LC-MS analysis?

    LC-MS, fungal metabolities

    Muthuramkumar Sankaralingam · Virudhunagar Hindu Nadars' Senthikumara Nadar College

     Thanks for your suggestions

  • Arpit Bhargava added an answer:
    How can I remove fungal spores from extracellular fungal protein sample?

    I am working on fungal extracellular proteins. In near future I would be separating various extracelluar protein by gel filtration chromatography (GE healthcare AKTA PURE FPLC system) using Superdex 200 increase column. I am worried that presence of fungal spores in my protein sample may cause problem as these spores may grow within the column thus making it impossible to use the column. Can anyone suggest me how to completely remove fungal spores from protein samples?

    Arpit Bhargava · Birla Institute of Technology and Science Pilani

    Respected Tarafdar Sir .....Thank you for your suggestion.... 

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