looks like you may have contaminated the coconut at 5:04 when you put the scalpel inside the flask where your fingers had been... maybe that is why it didn't perform as well...
Ryan The was no contamination. Every thing is sterilizing my hands get wash and sterilize with alcohol. Also I checked the 3 cultures in the microscope and found no bacteria. I did not put that in the video to keep it short. Actually toward the last day it got a bit better.
Hi Carlos I'm Enrico from Italy, nice job and propagation setup, I liked the video a lot. Can you please tell me the antibiotic agar recipe you used to isolate beauveria? I'm "playing" with a jar with some infested stinkbugs lately, they do lots of damage here, I add new ones to the jar and they're gone in very few days, mycelium grows outside them and sporulates. In another jar I used my compost and an inoculum that "could contain" spores and I'm seeing beauveria and something similar to metharzium killing stinkbugs and other pest. I'd love to isolate them on agar first and then making pure liquid cultures, but I'm having contamination issues with a simple nutrient agar. I'd also like to address you to a paper about the production and efficacy of beauveria bassiana blastospores produced under liquid fermentation in liquid cultures, different from the conidia on the surface mycelium mat you are producing, I think you'll find it interesting, try to search it on google. Do you think your method produce more conidia than a jar of sterilized rice (solid state fermentation)? I'm thinking about the surface volume could be a lot more I saw you're using beauveria in drench applications, are you trying to give them as plant endophytes? I heard they can live inside plants. I grow mushrooms in autumn/spring and vegetables for my family all year round and I'd also like to use entomopathogenic fungi as foliar spray at certain times of the year. I'm in a facebook group about soil microbes and biofertilizer production if you're interested, lots of good material there: some people would love to see your video, know your agar recipe and ask you something, so I'm going to share it. We were discussing this argument last week. Thank you in advance and have a nice day.
A lot of questions: The jar with contaminated bugs with Beauveria is a good place to start. I don't do Facebook you will find that Tweeter X is a better way to communicate for Scientific work. My handle @carlosdlt280 Selective media for beauveria bassiana is a little more complicated to make I have used some with Dodine, hard to find and expensive. YOu can ask Chat GPT for selective medias to isolate Beauvera and would give you good choices. This other does not require Dodine The OA-CTAB medium is composed of: • Oatmeal 20g/l, • CTAB (is heat stabile - add before autoclaving) 0.6 g/l, • chloramphenicol (is heat stabile - add before autoclaving) 0.5 g/l and • agar-agar 20 g/l. Colonies are smaller and sporulate faster than on dodine. it takes more incubation time. approximately 5 days at 27ºC. Cook the oatmeal and strained it through cheesecloth. Chloramphenicol and CTAB can be added before autoclaving as both seem to be heat stabile. Dra. Julieta B. Posadas Laboratorio de Hongos Entomopatógenos Instituto de Microbiología y Zoología Agrícola However, in your case I would make a Potato Dextrose Agar or Sabouraud Dextrose Agar add some antibiotic and pick a very small piece of the mycelium from the bugs and do several plates then isolate from those to end up with a cleaner culture. May try with or without antibiotics. After that you can isolate it mix with 20% Glycerol and store in the refrigerator to be used in the future. ChatGPT is very good at this. Good luck
its appreciated, well-done, also looking for application and response of the product
How long and under what conditions can beuaveria and trichoderma be stored?
@@adammatuszewski9822 there are many ways to store. I suggest you ask Chat GPT AND NARROW it down to what is available to you.
How do you know how much solution to use for what amount of water? In bauveria and trichoderma?
I put the recipe in detail on the film. I dont grow Trichoderma in liquid i do it in grain
@ yes i know i watched your video with Trichoderma but why u do it on the grain? Its better way to produce?
@@adammatuszewski9822 yes is better aclotcmore productive
@@GraftingAvocados Thank you very much
What are you trying to control?
@@MilesTaylor-s7p Basically testing metabolites of some commercial bio controls vs Laurel Wilt pathogen
looks like you may have contaminated the coconut at 5:04 when you put the scalpel inside the flask where your fingers had been... maybe that is why it didn't perform as well...
Ryan The was no contamination. Every thing is sterilizing my hands get wash and sterilize with alcohol. Also I checked the 3 cultures in the microscope and found no bacteria. I did not put that in the video to keep it short. Actually toward the last day it got a bit better.
so 1mL=10 CFU?
@@robertcepek5261 if you mean total cfu per ml from the concentrated solution of the 3 flasks. From prior work I have done is at least 100000 per ml
seems you leant from mr sunil kumars video well :)
Hi Carlos I'm Enrico from Italy, nice job and propagation setup, I liked the video a lot.
Can you please tell me the antibiotic agar recipe you used to isolate beauveria? I'm "playing" with a jar with some infested stinkbugs lately, they do lots of damage here, I add new ones to the jar and they're gone in very few days, mycelium grows outside them and sporulates. In another jar I used my compost and an inoculum that "could contain" spores and I'm seeing beauveria and something similar to metharzium killing stinkbugs and other pest. I'd love to isolate them on agar first and then making pure liquid cultures, but I'm having contamination issues with a simple nutrient agar.
I'd also like to address you to a paper about the production and efficacy of beauveria bassiana blastospores produced under liquid fermentation in liquid cultures, different from the conidia on the surface mycelium mat you are producing, I think you'll find it interesting, try to search it on google.
Do you think your method produce more conidia than a jar of sterilized rice (solid state fermentation)? I'm thinking about the surface volume could be a lot more
I saw you're using beauveria in drench applications, are you trying to give them as plant endophytes? I heard they can live inside plants.
I grow mushrooms in autumn/spring and vegetables for my family all year round and I'd also like to use entomopathogenic fungi as foliar spray at certain times of the year. I'm in a facebook group about soil microbes and biofertilizer production if you're interested, lots of good material there: some people would love to see your video, know your agar recipe and ask you something, so I'm going to share it. We were discussing this argument last week.
Thank you in advance and have a nice day.
A lot of questions: The jar with contaminated bugs with Beauveria is a good place to start. I don't do Facebook you will find that Tweeter X is a better way to communicate for Scientific work. My handle @carlosdlt280 Selective media for beauveria bassiana is a little more complicated to make I have used some with Dodine, hard to find and expensive. YOu can ask Chat GPT for selective medias to isolate Beauvera and would give you good choices. This other does not require Dodine
The OA-CTAB medium is composed of:
• Oatmeal 20g/l,
• CTAB (is heat stabile - add before autoclaving) 0.6 g/l,
• chloramphenicol (is heat stabile - add before autoclaving) 0.5 g/l and
• agar-agar 20 g/l.
Colonies are smaller and sporulate faster than on dodine. it takes more incubation time. approximately 5 days at 27ºC.
Cook the oatmeal and strained it through cheesecloth. Chloramphenicol and CTAB can be added before autoclaving as both seem to be heat stabile.
Dra. Julieta B. Posadas Laboratorio de Hongos Entomopatógenos Instituto de Microbiología y Zoología Agrícola
However, in your case I would make a Potato Dextrose Agar or Sabouraud Dextrose Agar add some antibiotic and pick a very small piece of the mycelium from the bugs and do several plates then isolate from those to end up with a cleaner culture. May try with or without antibiotics. After that you can isolate it mix with 20% Glycerol and store in the refrigerator to be used in the future. ChatGPT is very good at this.
Good luck
Gas🎉