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Thanks! I'm glad you found it helpful, I know this can be a complicated topic. Best of luck with your studies.

Oh thank you, I'm glad you found it helpful!

Glad it's helpful!

Good luck with your exam! I hope these videos are helpful for you.

You're very welcome!

First of all, good luck with your dog's treatment! Aspergillus can certainly cause respiratory infections in dogs; based on the diagnostic tests the veterinarians at WSU performed, I suspect this is what they identified. A BAL would be common test used to identify the presence of infectious agents in the lung. It certainly sounds positive that this infection is isolated to the lungs, systemic aspergillosis is a very serious and difficult to treat infection that most commonly occurs in dogs which are immunosuppressed (either due to treatment with an immunosuppressive drug, or because the animal has what we call a primary immunodeficiency). As a microbiologist, I don't personally see patients so I don't have first hand experience with treating these types of infections. I know that there are some excellent specialists at WSU, so I suspect your dog is in very good hands!

@@therubinlab Thank you! I appreciate it.

You are most welcome!

Good question, and I think that's a common misconception. I don't think I would say that this means that the treatment is weak - resistance means that the drug is not predicted to work if used according to it's label indication (in the species intended, dosing regimen - dose, route of administration and duration, and for the type of infection it's labeled for). I've put together another video which you might also find helpful, there is also a link to a written/visual guide which you can download: ruclips.net/video/RRFlSIKVSEs/видео.html I hope you find this helpful!

Yes! We always incubate plates upside down, otherwise you can get condensation on the lid which drips down onto the agar surface. When doing disc testing, it's important to gently push on the disc after it's places so that the discs don't fall off.

@@therubinlab thanks for your prompt response. I think it’s really important to mention this step in your video so the method will be clear to the end.

That's a really good point... I'm not sure if you saw, but I recently published a video which includes a link to a guide to performing disc diffusion testing ruclips.net/video/RRFlSIKVSEs/видео.html My co-authors and I tried to make this guide as clear as possible... have a look and let me know what you think. I hope this resource will be helpful as well!

I'm so glad you've found it useful! Best of luck with you studies :)

You're welcome!

Sorry for the slow reply! For a study with the aim of generating some preliminary results, I would say that it's less critical which kind of media you use. It might be more important to consider the properties of the compounds that you're hoping to test (what is it soluble in? how diffusible is it?). When working with a novel compound or a complex substance like a plant extract, I would suggest performing MIC testing - I previously made a video describing the agar dilution method that you might find helpful: ruclips.net/video/1t_UQhtR7pg/видео.html Best of luck with your research!

Being in the same family these drugs would have similar properties (same mechanism of action, and similar spectrum of activity), but since these aren't drugs that we use in veterinary medicine I'm not as familiar with the other examples you mentioned. Sorry I can't provide more details on these compounds!

You are most welcome! I'm glad you're finding them helpful :)

Oh you are most welcome, I hope you find it helpful! Good luck with your exam :)

I'm really sorry to hear about your kitten! Unfortunately, because there are so many factors that impact treatment effectiveness and every animal's situation is different, it's not possible to suggest alternate therapies. Actually, as a veterinarian I am not legally allowed to provide advice without seeing your kitten as a patient so that I can understand his or her particular situation! One thing that I could suggest, is that if your kitten's situation is particularly complicated, you could request that your family vet refer you to a veterinary specialist for a more in-depth workup. Best of luck, I hope your kitten gets better soon!

i’m learning micro for the clinical setting right now and this was full of fun facts outside of “clinical significance” even thou they are why it’s significant lol. thank you

@@oscrates I'm glad you found it useful! I tried to include a good mix of clinical significance, and all the background information necessary to critically evaluate infectious disease cases. Best of luck with your studies!

The first step in solving any recurrent health issue in an animal is to ensure that you have an accurate diagnosis, that way your veterinarian can treat the root cause of the problems your cat is experiencing. So I think the only advice that I can really give you isn't about which drug to treat with, but to suggest that if your family vet hasn't been able to adequately address your cats health problems, to perhaps request a referral to an veterinary internal medicine specialist if you have one in your area. Sorry, I can't give you any specific advice about your cat, best of luck!

@@therubinlab we mad all test and laboratory tests said in culture its M.staph intermedius According to that wrote the sensitive and resistant antibiotics we used amikacin and now the doxycycline which are wrote as sensitive to staph but the result same iam very tired what to do

@@therubinlab all test are normal But they advised me to continue more one week with doxycycline is it right or should i change it to another

I'm sorry about all the frustrations you're having, one other thing that I forgot to mention is that it's not possible to simply recommend a medication for your pet based solely on the bacteria which was isolated. Each animal's situation is different, and so while drug X may be the best option for one animal, it might be a bad choice for another infected with the same organism - so there is unfortunately no one solution that could be recommended. Best of luck with your cat!

Glad it was helpful, good luck with your studies :)

I hope you found this description of Mycoplasma infections helpful!

Thanks for the comment! I just recently uploaded a new video, and step-by-step guide you can download, about disc diffusion testing - you can find it here: ruclips.net/video/RRFlSIKVSEs/видео.html Good luck with your studies!

Oh thank you so much for the comment! The reason for the diamond shaped lesions is the geometry of the vascular anatomy... in part 1 of this lecture I briefly describe the pathogenesis; here's a link to the explanation: ruclips.net/video/jD7xmXohiA0/видео.htmlsi=-Xmyiu3A0-V_Tyiw&t=491 Hopefully that answers your question!

@@therubinlab 💙 thank you for your help!!!

My pleasure! It's a great collaboration.

You're welcome!

You are most welcome! I hope having a copy of the notes will be helpful

Hi Jamie - sounds like an interesting project. I might be misinterpreting your question, but think there might be a bit of confusion with the definition of MIC - this is the minimum inhibitory concentration; in order to determine the MIC you have to test multiple concentrations. If for instance you test 1, 2, 4, 8, and 16 micrograms/ml and you see growth of an organism at 1, 2 and 4 micrograms/ml but not 8 or 16, then the MIC would be 8 because this is the lowest concentration which inhibits growth. I hope that makes sense and helps! If I've misinterpreted your question please just write back.

Glad you liked it!

Good question. I'll try and answer in point form for clarity: 1. Yes, you could use agar dilution for the plant extract. You just have to be able to dissolve it into a liquid form prior to incorporating it into agar. Depending on how much of the extract you have, it might be more practical to use a broth-micro dilution method (since plate volumes are 20ml you will likely need to use a lot of the extract). 2. The McFarland standard is to make sure you're testing a consistent, standardized concentration of bacteria (although with fungi there may be a different method). I'm not familiar with anti-fungal susceptibility testing, I would suggest looking into the CLSI guidelines to see what the standard procedure is. 3. For this study, I think it will be really important to also include a positive control (no extract plate) to ensure that your fungus will grow on the media you use. Again, here I'm not sure what's recommended for fungi, Mueller Hinton is the media used for non-fastidious bacteria. The M27 and M38 documents describe broth dilution methods of anti-fungal susceptibility testing (so broth might be the way to go rather than agar!) I hope that helps, good luck with your experiments!

@@therubinlab Thanks sir for replying! We’re not really sure if broth-micro dilution will be feasible in our part since our university still lacks some of the equipment needed. Do you have any other methods that you can recommend for testing anti-fungal activity specifically by using a plant extract? I’ve been trying to research about it but the results are so minimal.

@@kelseymallow8428 I think the best approaches would all be based on determining an MIC, and I think either broth or agar dilution would be the way to go. For the broth method, you could also do "macro-dilution"; here instead of using commercially prepared 96 well plates, you prepare broths with doubling dilutions of your compound in larger tubes (maybe 2-3ml volumes). This should eliminate the need for any specialized equipment (just tubes, pipettes, incubators etc.). If you can get your hands on the M27 document (clsi.org/standards/products/microbiology/documents/m27/) you should find some detailed instructions on how to perform this test.

@@therubinlab Is it possible to use agar well diffusion method? It’s the most feasible method in our part. How is the MIC usually measured if this is the method used? In terms of making the dry plant extract into a liquid, can we use distilled water for dissolving the dry extract? Thank you much, your replies are really a big help in our research.

@@kelseymallow8428 I'll try and answer in point form for clarity: 1. I'm really not sure about the appropriateness of agar dilution vs. broth dilution for anti-fungal testing, unfortunately this just isn't an area that I have expertise in! I'm not familiar with the agar well diffusion method; is this putting extract in a well in the plate and looking for a zone of inhibition caused by diffusing anti fungal compounds? If so, I think this sounds like a reasonable approach but would be very qualitative (I think the only question you could reliably answer is - does extract X have any impact on the growth of the fungus we tested). Further studies would be required to quantify the inhibitory effects. 2. In agar dilution, the MIC is defined as the lowest concentration which inhibits the growth of the organism - so if you have growth on the plate with 2ug/ml, but not 4ug/ml then then MIC would be 4. 3. Distilled water may be an appropriate solvent for the extract, but it really depends on the chemical properties of the substance present. When preparing antibiotic suspensions for agar dilution we do use water for most drugs, but there are some where we first need to solubilize the powder with ethanol, DMSO or PBS and then dilute the solubilized compound with water. I hope this is helpful; I appreciate that there are a lot of complications!

I'm glad it's helpful! In the next week or so I am planning to make all my lecture notes available, I'll put a link in the upcoming video :)

Oh you are welcome! I'm glad you found it helpful :)

Good question! Yes, there is a specific medium - both the CLSI and EUCAST state that you need to use Mueller Hinton agar. Nutrient agar does NOT give zone diameters which you can interpret with recognized criteria for classifying an isolate as susceptible or resistant. It's also important to make sure that plates are 4mm thick so that the drug diffuses through the media in a predictable way. Stay tuned to the channel... I just filmed demonstrations for a new video and took photos for a new visual guid that I'm preparing - I'm anticipating that video should be out in the next 3-4 weeks :) In the mean time, if you have any questions please just leave another comment below!

@@therubinlab Thank you! Much appreciated!

Oh thank you for the feedback, I'm glad you're finding them useful :)

Glad you've found it helpful! Great to know that these lectures are useful for students in Uganda as well as my own classroom :)

Perfect! I'm so glad you found it helpful :)

Thanks, I'm so glad you're finding it helpful! I won't be doing any virology videos, but I am planning on making pdf copies of all my bacteriology lectures available in the next little while... stay tuned for a new video which will contain a link to those files :)

Most welcome 😊

I would love to come to Mexico! There are so many interesting organisms that I would love to come and learn from your experts about... especially Sporothrix and Coccidioides, bugs we just don't have in Western Canada. So much to learn from each other!

Thank you so much for sharing the videos with your colleagues! I couldn't be happier than to have these videos reach more students :)