All right, dermatology sampling. This is when we actually get to put our hands on the patient and find out what's going on. So our first sample that we would take would be a coat brushing because this is quite easy to take.
You can use a flea comb or you can just ruffle the coat a whole lot with your hand like you give them. Back a good rub with your hand and you'll get lots of Coat, hair and little bits of scurf and stuff like that falling out onto a paper towel or with the brushing, you can just pull it off your calm. This is done to check for parasites.
I'm sorry about the blurriness, but my, little neotrambiculo, when I expanded him, he did look a little bit blurry, but that is a harvest mite in the larval stage, which is why it only has 6, not 8 legs. . So you could catch a harvest mite with a coat brushing, although, I usually get a better job doing tape strips with those guys.
You can get Kayla Tello with coat brushings. Good way to collect samples for dermatophyte testing. So you might do your coat brushings, put them in a universal pot, and send that off to the lab.
Or, what I like to do if I'm doing an in-house dermatology test, dermatophyte test is get a toothbrush. From the supermarket, you get them from a lab, but they're a little bit more expensive if you buy them from a lab. If you get them from the supermarket, they're, still sterile until you open the pack, and then you brush the animal around the lesions or if it's a cat everywhere, and then you embed the toothbrush into your agar plate, and that's a really good way to grow some spores if you suspect amount of phytosis.
Cut brusings help you assess the level of surf, help you identify any flea dirt, and it's a good way to collect samples for trichography if you're gonna look at, hair shafts to see if they're in the telage stage, the late stage when they're not really growing any longer, or if they're shattered or have follicular debris. Tape stripping, I love tape stripping because it's you're just taking a piece of stationery and then you're doing science with it. So, Kaylee Tella, is walking down drift.
There's a little one there. He's pulled his legs in, so it's kind of hard to tell. He's got, eight legs, but you can see the little biting pallops at the front of the, the.
Well, on his head, basically, he's got little pinch, biting nasty little pincers, hard little guys, and they walk around in the, on the surface of the skin and chew on scurf and stuff like that, and that's why we call them walking down drift. Supposedly you can tell it's a surface mic cause it has longer legs, but obviously in this photo, he has tucked his legs under him. And they're supposed to have a a nice trim body cause they do so much exercise walking around, whereas a sarcoptes mite is a big fat round mite cause he just sits and burrows and, and it eats things, and he doesn't do as much exercise as aylo ella.
Whether that's a myths or not, I don't know. Neo trabecular autumaliss is the, larval stage of the harvest mite. This looks like little red dots, and you can catch those with, a tape strip.
They like hanging out in the Harvey's pocket, the Henry's pocket, somebody's pocket in the ear, that little fold in the ear, you see them there and you can get them between the toes. Sometimes you'll get a cat that has basically ripped its face off. And you're like, oh my God, what's wrong?
Do some tape strips, you find the harvest mite. Rub the poor little kitty with a little, you spray some fipronil on some cotton wool, rub it, kill the mites. You fixed it like that.
For some reason, kitties get a real, some of them get really hypersensitive to harvest mite. So if you ever see a cat that has really, really hurt its own face, definitely check out for harvest mite, especially if you live in an area with chalky downs, and it's harvest time. Cytology is also done with the tape strip, so you would apply the tape strip to the affected area.
And then put a drop of methylene blue or or diff quick stains to identify what you're looking for. So cytology or looking for bacteria or yeast and it can be found, this simple piece of office material. Flux, I like using flux for Damodex, .
People do a lot of people do scrapes for Damodex and you can catch them that way, but I've just been a lot more successful with plucks. Once I accidentally plucked a whisker, I'm really sorry, dog, I didn't mean to, but I caught 17 mites on that one whisker. So it's a real effective way of finding them.
Also, if you've got lesions around the eyes, it's nicer to be plucking rather than scraping, so. Also feet, don't like scraping feet. No, no.
Definitely better to pluck. So I use artery forceps, usually the ones that the vets say these are crap. I'm not using them anymore.
So I take those out of the kits and I use them. Some people use tweezers. I don't like tweezers, they're too sharp and pointy.
Also useful plucks can be very useful for your dermatophyte testing. So if you want to throw these on your agri plate or put them in a universal pot and send them off to the lab, good way. And again, looking at your hair shafts, are they fractured?
Are they old tillage and hairs? Are they covered with follicular debris? All these things can tell you things that the animal it's chewing on its fur, or it might have demodex if it's got lots of follicular debris or seporrhea, .
Or lots of teenage and hairs would indicate maybe hyperthyroidism. Skin scrapes, this is, to me, this is the nastiest of the samples that we do. What you need to do is trim some hair around.
So I don't like using clippers because you could actually catch, you know, shave off a mite that you need. So just use scissors to shave away any hair. Squeeze the skin.
And then scrape with a number 10 blade until you see capillary ooze, so you must scrape until the animal bleeds. So this isn't nice, and you need to tell the owner beforehand, so they're, you know, you're not turning back a dog or a cat with big Sores all over it. You say, I'm gonna need to take at least 5 samples from your pet, and those will look like red sore spots afterwards.
I will apply some soothing cream so you can apply whatever you choose to use in your practise, and just let them know ahead of time and say, I'm really sorry that this is necessary for the diagnosis, . So they're not going to get really angry and make a complaint letter. But yeah, this is not, this is one of those things where you've got to wait to have the examples for the student nurse to learn it because they can't just practise on any dog.
This is a little bit painful and we only do it if we really feel like we have to eliminate the possibility of mites. And this would be done for sarcoptes GBI or Demodex. And that is a Poor little puppy there with a really, really bad infestation of sarcoptes.
Someone should have done a skin scrape a long time before the dog got to that state. Impression smears? You can just take a good old slide.
So this is kind of your tape strip can do this or your slide can do this. So you just press the slide right onto the lesion. This dog here, the nose, those are really good lesions there.
Oh, they're ruptured pustules, so you're gonna find all kinds of interesting stuff on that slide. And that's a nice place where you, if you, you, you're not gonna scrape there. Oh, no, that would be horrible.
So you just press a nice little slide, you know, really calmly telling the dog that he's good, . And get your sample. I even managed to do this to my whippet, who is scared of life himself.
I mean, he spends his entire life hiding under a duvet, and he did tolerate having a slide pressed to his neck without having a complete mental breakdown. So this is very non-invasive and it's very good for lesions. And if you want to stain it and look for bacteria or yeast, tumours, you might pick up tumour cells this way.
I'm not great at identifying tumour cells. I can say, yeah, those look like tumour cells. No idea what kind of tumour.
That's not really your Job, but you can say to the vet, this is worth sending off to the lab. This is definitely worth biopsying because this isn't just a bacterial lesion, there's something else growing here. So that's quite helpful and ears, you can do impression smears with ears.
I call this cotton bud cytology. That's not a technical term. I think I've made that up.
Basically, you just stick a cotton bud into the lesion or into the ear and roll it on a slide. And with ears, I like to do a little L for the left ear and a little R for the right ear. So I'm rolling the slide in the shape of an L and an R.
And then when I stain them, I can go, oh yeah, this was the left ear and this was the right, cause I might have forgotten to mark it. The reason you do that is you can grow very different things in each ear. You might have one ear growing cockeye and the other one growing malaysia.
You might have one ear full of everything and the other ear is clean as could be. So you need to know which was the left ear and which is the right. And if you do it with your, when you're actually applying your cytology, that's just bells and braces, but you should still write on the slide as well.
And then lesions, you can stick a a cotton bud into a lesion and, and find out, you know, is it gramme positive or gram-negative bacteria, if it's gram-negative bacteria, I need to send this to a lab for culture and sensitivity. Air is really important to find out if it's just normal, commensal bacteria that has overgrown in the ear. OK, we can probably treat it with Polypharmaceuticals, but if it's pseudomonas or some other gramme negative, then we need to identify exactly which bacteria it is and use the exact right antibiotics.
Otherwise we are not treating the animal properly, then we can contribute to antibiotic resistance, which is a no no these days. OK, preparing your samples for analysis. Generally speaking, your coat brushes, your plucks, and your scrapes.
You're gonna put it on the slide with a drop of liquid paraffin, or some other oil equivalent that you can use for slide preparation. I sometimes use the oil immersion that I get from the lab to prepare slides as long as you don't put it on the animal. Liquid paraffin you can put on the animal, but I know it's hard to get, so.
But preparing your slides, you can use the oil immersion oil. Place a cover slip on it. That's important because these mics, some of them are walking around, especially that, you know, Kayla Tella doing her, you know, getting it 10,000 steps a day.
So you got to put your cover slip on or your mic might walk off the slide, and then you're gonna view it on times 4 or times 10. Some coat Russians, yes, you are going to put into an agar plate for it to mattify culture or send to the lab for your tomatify culture. That's a different story.
Tape strips. When you're doing them for parasites, you just do your tape strip on the animal and then stick the tapes directly to the slide, and it's its own cover slip. The tape is its own cover slip and you can just look at it like that.
If you're doing cytology, say, ear cytology, for some reason that's the one that has to always be done in a hurry. I guess it's because they want to decide whether they need to do culture and sensitivity before the animals at home. So that one the quickie way to do it is tape your, put the tape on the ear.
Put one drop of methylene blue or the blue in your cryptiff stone, the dark blue, and then press down the slide and you can look at it right away using the tape as your cover slip. So you don't have to. Dry it.
You don't have to multi-dip or anything like that. You're just one drop, press down the tape, and look at it. It's a really, really quick preparation if you're in a hurry to find out what's going on with that year.
You view on times 40 to see what's going on, and then move on to times 100 to look for your bacteria or your yeast. Another way to use your tape strip is to create a little loop at the end of the slide, and then you can dip the loop into your differential stains. Probably skip the fixative stain if you're doing an ear because that can sometimes dissolve things because the ear wax just dissolves in the acetate.
So, you can do that that way if you want to do it. It's a good, good way to do your cytology when you're not in a hurry is to use the differential stains. Impression smears and cotton bud cytology, again, you can stain with just one drop of methylene blue or you can dip in.
All of your 3 differential stains, plus or minus the fixative, if you're doing an ear, leave out the fixative. But if you're doing, say, an impression smear like on that, those horrible craters on that dog's nose, I would use all three on that one. Then you need to dry it very carefully.
You. Air dry is better. Air dryers are always better, but sometimes you don't have that luxury.
So, use a hairdryer on the low setting and only blow the back of the slide. Don't put any warm air on the front of the slide because you will deform the cells and then it won't be diagnostic. But if possible, do try to air dry.
And then you would examine times 100, which is oil immersion. Although I like to always find a good area of interest at times 41st, or even times 10, find a nice area of interest, and then go up to your times 100 because otherwise you might be just To scan the slide on 100 takes a long time, and you might be just going past really boring stuff for a really long time. So find your area of interest and then go back to that with your times 100 and do your .
Do your cytology report for the vet. I like, I have a little form I fill out and write, you know, from the foot, I found this from the ear, I got this, and I give the written report to the vet and then they can diagnose from that point because obviously nurses do not diagnose. Preparing your slides, I've got the methanine blue there as a separate because I use that a lot just on its own.
And then This is my cop cell pots here. You can see how messy it is. I've got an inco pad underneath them, so when I'm blotting my slides, it just goes into the inco pad and I change the inkopad every couple of days.
Do you have something to protect your work top cause these stains are pretty permanent, they're hard to get off. I've got two sets of cop cell slides. One is for blood and one is for dermatology.
Don't mix between the two cause you will get, you won't be able to use your slides. You will be so confused cause things that grow on ears don't grow in blood and it just won't make any sense whatsoever. So, separate the two.
Clean your pots at least once a month, at least. So all the stain goes down the drain, scrub the pot, get a good bottle brush, and really scrub because you want to get rid of that precipitate, soak them in, a, a cold disinfectant. Bath, probably something that you use for some of your surgical instruments.
Soak them there for 10 minutes, then rinse, rinse, rinse, then put your new stain in. I know that sounds really labour intensive, but otherwise, you could misdiagnose things cause you could have dipped a slide that had loads of bacteria. Some of the bacteria slid off into the stain and you're sharing it.
With another patient and you're gonna say, oh, this ear is all full of bacteria, when actually it was another animal's ear. I prefer at this point to use pipettes rather than actually dipping the slide in the stain because it it limits cross-contamination. So if you can possibly use pipettes instead of dipping your slide, it will keep your stains a lot more clean and your diagnosis more accurate, your vet's diagnosis more accurate.
Yes, this is a waste of plastic, and it really hurts me that I'm doing that. So what I try to do is I set out a pipette for each colour stain and use the same pipette all day, so I'm not throwing away plastic pipettes, you know, every 5 minutes. That is OK.
You're not going to, commit cross-contamination that way. So You know, that's one way to do it. So there's an example here of pipetting the the stain on and then pressing down the slide with the tape strip.
So you can then examine it under x 40 and times 100. A little examples here, these on the left side are stained slides. One was an impression smear and one was a tape strip.
And then you've got a tape strip that was not stained. This was just directly taken from the coat and the tape pressed down. So, the plain one is parasites, the Dark one on the bottom, I'm saying that's an ear and then the impression smear, that would have been from a lesion somewhere on the animal.
And here you have your lovely microscope. I find a lot of people are really afraid of their microscope. I don't know why.
I don't know what happens to you at school, that makes you afraid of microscopes. Maybe the teachers yell at you, be careful, you're gonna break it. I don't know, but they are pretty sturdy.
Don't be afraid of them, make friends with them, get used to all the moving parts, . If you need to check back on your old vet nursing textbook on a diagram on the microscope, that's fine. Nobody needs to know that you forgot about the parts, but What I say to student nurses is if you spend, if you get a lunch break, I know a lot of nurses don't, but if you get a lunch break, take 5 minutes of your lunch break or 5 minutes at the end of the day, every day for a month and just Look at things under the microscope.
No one's breathing down your neck. Nobody needs the information immediately. Nobody's gonna yell at you if you've got it wrong.
Just look at things. At the end of the month, you will be so comfortable with the microscope. The microscope will be your best friend, seriously.
So, get used to it. Learn how to move the stage up and down. Learn how to move the turrets, which turrets work best for, you know, the sample you're looking at.
Get used to the eye pieces. Are you happier with one? Do you want two?
Yes, they'll make you dizzy at the beginning, but you will get used to it. Figure out where the rheostat is, you can lower the light intensity, . Figure out where the iris is.
Remember the Vernier scale if you wanna go back to a specific spot on the slide. It's all stuff you learned. It's just triggering the memory, and it will make you a much better nurse in a lot of ways cause you can look at blood films, you can look at urine, it will make you very valuable.
At my current practise. I look at just about everything under this microscope and they're always going, can you do this, can you do that? And if you become good at it, you'll be so valued by your practise.
So definitely worth the effort. I have this posted by a microscope because I don't like it when people are mean to my microscope. So these rules are important to remember.
There is a skin product company that will send you a nice little poster about how to care for a microscope that you can put up next to your microscope if you want, but I made my own. Turn the ristor all the way down. And make sure your stage is at the lowest height before you turn it on.
So the start on some microscopes is like a little dial on the side, basically it just turns the light levels all the way down. You're gonna use x 4 x 10 for parasites, x 440 for urine or blood smears, and 100 is for bacteria or yeast. When using the 100, don't pour immersion oil on there.
One drop will do a lot. When you are done, make sure you clean the lens. Don't ever use leave the oil on the lens because it will stick there and get gummy, and then you've lost that lens.
It's no good. Make sure that you never, ever, ever get oil on any other lands, and it seems to be the times 40, people keep, they go back and forth between the 40 and the 100, and then they ruin their times 40, and then you have to get someone to come in and professionally clean your microscope. So just be really conscious of which turret you're using and which lens and make sure that you don't mix them up.
You should clean every lens. What I do is I get one lens tissue out, I clean every lens, and then the times 100 gets its own special tissue, so they don't mix up like I don't want to clean the 100 and then use the same tissue for the x 40 because then again, it will be a problem. Yeah, and be considerate and clean up after yourself.
These are biological samples and they can, spread disease if we, if they're not cleared away. And do make sure you have someone come in and maintain your microscope, clean out all the dust, and, clean your lenses every 6 to 12 months. There are a lot of companies that will come to your practise and do that, so just look them up and book an appointment.
These are photographs of of slides that I have made. And took the picture with great difficulty. I'm not very good at that.
But the first slide all the way on the left hand side of the screen is malashesia. So those are yeast. They are often found in the ear, sometimes they're found between the toes, and they can be found on skin.
Malaysia is a commensal, so it lives on the body and it's very happy there, but it overgrows when there is some kind of immune problem or a, Trauma to the skin. So you will see it in ear infections. It's normal to see 1 malashesia per high power field.
So high power field is times 100. So if you see one malace it's, ah, no big deal. So if you see 1 per field, like you look at 10 fields and you've seen 1 or 2 per field, you're not going to get too excited about it.
To see this many in a field is very bad. That year is got a terrible mala easy infestation, and it really, really, really needs some attention. If you saw that on the skin and you decided to prescribe antibiotics, that would be silly because a yeast isn't a bacteria.
You need to use something for that will kill yeast, if you've got yeast growing. So it's always worth finding out, doing your, your tapes strips or your impression smears and finding out what's growing on the skin. Generally speaking, we can use something nice and topical, and some of the topical treatments will do bacteria and yeast at the same time, but if it's a deep, skin infection, And the vet feels that a systemic treatment is necessary, make sure that you're treating it appropriately and not giving antibiotics to something that has a yeast infection because it will be a waste of time and money and it will cause antibiotic resistance.
The middle slide, you can see some neutrophils there, and if you look carefully, you can see some, rods. Those are gram-negative bacteria. That is definitely something where you need to do a culture and sensitivity to make sure you're using the correct, Medication to treat that because if you don't, again, you're gonna get antibiotic resistance, but more importantly, you're not gonna cure the problem and the animal is gonna continue to suffer.
So this is Probably also from an ear, and it might be, it looks like it's probably pseudomonas. So definitely this is if you see this, then you're saying, oh, we're gonna have to do a culture, send off a swab. In the meantime, let's, you know, keep the animals comfortable as possible, but we need to maybe wait on deciding which treatment to use because we don't want to give the wrong thing.
And the other slide is a gliella, and that's a surface smart. This was done with a tape strip. You can, a lot of people mix them up with oddecties, but oddecties have little suckers on their feet, and with Glet you can see really nasty little biting mouth parts as well.
