This action might not be possible to undo. Are you sure you want to continue?
Dermatology Diagnostic Procedures
Confirms presence of surface dwelling ectoparasites Parasites that can be identified in this way are: - Fleas and flea faeces - Cheyletiella (mites) - Lice Technique: 1. Place px on floor standing on white or brown piece of packing paper. 2. Vigorously rub skin and haircoat to help dislodge material that may be adherent. 3. Follow by combing vigorously with a flea comb including the entire body surface. 4. Move px away from paper and fold paper so all material in middle. 5. Remove the hair- parasites likely to be amongst the smaller debris. 6. Use strips of clear adhesive tape to remove remaining material from central part of paper. 7. Place on microscope slide + examine.
Rapid way of gaining important info about a skin disorder. Indicated in most cases of skin dz and in ALL cases of otitis externa. Stains used: Diff Quik = modified Wright’s stain (Romanowski). Methylene blue may also be used. Diff Quik = Alcohol fixative (clear) Eosinophillic xanthene dye (red) Basophilic thiazine dye (blue) Distilled water. Permits visualisation of bacteria, although sometimes a gram stain may be helpful as well. If mycobacteria are suspected an acid fast stain should be employed. Technique: Impression Smear 1. Press a microscope slide onto skin (clip hair away) or lesions/pustules if the epidermis is eroded. This can also be used for sampling ruptured pustules and the cut surface of tumours biopsied or removed, (squeeze +express the contents onto the slide). 2. This is useful for assessing levels of Malassezia (yeast organisms naturally present on skin surface but if overgrowth occurs= dermatitis +intense pruritis) or bacteria on the skin surface. 3. Also useful for open, exudative, or draining lesions. 4. Be sure that the material is not too thick on the slide-smear it with the aid of another slide if necessary. 5. Demodex can often be found in this way from lesions of associated deep pyoderma.
One end of the tape is then freed and the tape pressed down onto the surface of the microscope slide. .6. Additional material is then obtained by an impression smear placing a microscope slide onto the ruptured lesion. using another microscope slide. Press firmly on area of skin or lesion to be sampled. as this may remove some of the lipid in which the yeast is generally situated – and with it some of the organisms. Before staining the slide should be air-dried and then alcohol fixed. The material is then air-dried and stained using the usual protocol. a coverslip applied. For Cheyletiella place straight onto slide +examine 1st under low power. if there is abundant material. When evaluating samples for Malassezia or ear samples heat is used. In assessing levels of Malassezia it may be preferable to omit the alcohol fixative stage of Diff Quik stain. Fig 1. A drop of immersion oil is placed upon the tape. Samples Procured with a Needle Using needle to rupture the lesion: A suitable lesion is gently ruptured using a 23-26g hypodermic needle Material from the needle tip is then placed on a microscope slide and smeared using the needle or. look for material from intracellular cocci. and the sample the examined under a high power and oil. Malassezia Tape Strips for Cytology Clear cellulose tape is used (sellotape). If examining material from suspected pyodermas. For Malassezia of bacteria each end of tape is then attached to the end of a microscope slide to facilitate handling for staining (allows dunking) Stain with Diff Quik omitting the fixative stage which would destroy the tape. 7.
If the mass were an epidermal cyst. If the mass was a lipoma. d. A cotton tipped applicator is introduced into the lesion After withdrawing material is transferred to a microscope slide by gently rolling the tip on the slide rather than smearing it. heat or alcohol fixation is undertaken prior to staining. c. liquid paraffin prior to introducing. It is also used for obtaining samples from ears. Use 20g needle or 18g if sampling more dense lesions. Partly withdraw and redirect 4-5 times to obtain additional material. Sterilise the surface of the mass with a suitable antiseptic. After application of a suitable antiseptic to sterilise the surface the needle is introduced into the lesion at a 30° angle to the skin surface. Withdraw the needle. Cytological Examination of Exudate from the Ear Canal A cotton tipped applicator is introduced gently to the level of the horizontal canal.FNA: For obtaining material from cysts or deep inflammatory lesions. b. Withdraw the needle. g. e. dry. routine expression of epidermal cysts in this way is not recommended. granular cheesy material that is amorphous will be obtained and is also readily expressed from the puncture wound. d. Pre-tx with antihistamines is a wise precaution. f. A 20g needle is attached to a 3 or 5 ml syringe with the barrel pressed down. 2) Using a syringe and needle: a. The needle is introduced into the mass at 30° and suction is applied. The latter is used where preservation of cell morphology is important. Smear. e. If ear canal is dry or Otodectes suspected. stain. However. Be careful when sampling possible mast cell tumours as their manipulation can cause adverse reactions due to HISTAMINE release. b. Diagnostic cytology may be obtained for skin tumours but in many cases biopsies for histopathology are required to confirm the dx. Attach needle to 3ml syringe or 10ml if greater suction is required. or a cutaneous mass. the plunger of the syringe is withdrawn half way or more. lipid material only will appear on the slide. Material is gently smeared to avoid too thick a preparation using either the needle or another slide. c. f. Air dry and stain. . moisten tip w. the needle is attached and the contents of the needle expressed onto the surface of a microscope slide. Air-dry. Don’t let the needle penetrate the deep surface of a potentially malignant neoplasm as it could aid in its spread. Reattach the needle and express the contents onto a slide. Gently rotate the tip in a circular motion to obtain the exudate then smear onto microscope slide. 2 main approaches for FNA: 1) Using a needle: a. Samples Procured with a Cotton Swab Indicated for sampling fistulous tracts and deep draining lesions. remove the syringe and withdraw the plunger.
50g chloral hydrate . which may be attached to the hair shaft.Telogen hairs have spear-shaped heads Telogen - Anagen Assessment of Anagen/Telogen ratios is not generally helpful as normal animals have widely ranging ratios. moisten the ear canal prior to sampling.25ml liquid phenol . For examination of bacteria and yeasts. Hair Plucking Useful where there is hair loss or abnormal appearance of the hairs. placed on oil.25ml lactic acid Both KOH and Chlorphenolac are CAUSTIC. (A2) . dependant upon the stage of the hair cycle.In looking for parasites additional oil is placed onto the slide which is then cover-slipped prior to microscope examination.Look for evidence of Demodex mites which may accompany epilation of the hair from the hair follicle.caution! Findings: The nature of the hair bulb . Evidence of parasites . (A1) .Look for anagen and telogen hairs . Technique: Sample of hairs grasped either with eyebrow forceps or haemostat.Look for evidence of eggs of Cheyletiella. and examined microscopically first under low power. Fix slide by heat rather than by alcohol then stain. Transferred to slide. (A3) - A1 A2 .Look for the ‘nits’ of lice.Anagen hair bulbs have rounded and smooth ends that are usually bent . If a dermatophyte is suspected a sample is also placed either in 10% potassium hydroxide and allowed to clear for 30 mins prior to examination OR in chlorphenolac which is .
The latter are seen in COLOUR DILUTION ALOPECIAS e. by excessive licking (Pic 2) . blue Dobermans (Pic 5).Examine the tips and ascertain whether they have pointed ends which is normal (Pic 1) . .A3 The nature of the hair shaft .g. e. which can suggest colour dilution conditions and excessive grooming.Look for evidence suggestive of trichorrhexia nodosa (Pic 8) which is a dystrophic condition of the hair or any other evidence of hair shaft dystrophy.g. Pic 2 Pic 1 Pic 4 Pic 3 Pic 5 Pic 6 Pic 7 .Look to see whether the melanin granules appear normal or if there are aggregates of melanin.macromelanosomes (Pic 4). and fawn Irish Setter and in black hair follicle dysplasia (Pic 7). blue Yorkshire Terriers (Pic 6).Assess whether the ends appear to have been sheared-off.Look to see if the hair shaft appears to have normal texture or of it appears more like a ‘ghost-shaft’ due to digestion by fungal keratinases.Look to see if there is evidence of fracture of the hair shaft. (Pic 3) . .
. Fungal hyphae(C) may also be seen. House dust mitesDermatophagoides farina. - A C B Intradermal Test Points to consider before deciding to set up allergy testing: How many pxs will you potentially skin test in a year? The outlay is expensive and the procedure is unlikely to be economically viable if used less than once per week. Dermatophyte arthrospores have a slightly greenish tinge(B) and a solid appearance that is appreciable upon focussing the microscope up and down. either in the hair shaft or in scale.Pic 8 Pic 9 Evidence of dermatophytes (Ringworm!) If you suspect dermatophycosis. Consult a veterinary dermatologist.should always be included. suggesting digestion by fungal keratinases. Carefully examine the borders under high power with a partially closed condenser (to find suspicious areas) and then oil immersion for confirmation. Choose hair shafts that appear as ‘ghosts’. Some 50% of cases of Microsporum canis will fluoresce green (A). first examine the haircoat with a Wood’s lamp. Human allergists usually use glycerinated extracts that are unsuitable for veterinary use. Which ones should I include? There is now good evidence on the incidence of positive skin tests in countries. Those of M. canis are smaller (1-2) than are those of Microsporum gypseum. Whose allergen extracts should you use? There are a number of reliable allergy houses that specialise and supply aqueous extracts. How many allergens should you use? There is no answer to this! Most use around 50 but it is acceptable to limit the screen to ~ 20 of the most common.
do not proceed). . parasitic and food allergy.g. In either case rpt the inj at an alt site. Should I purchase diluted allergens ready for use or allergens in concentrated form? Diluted allergens are only stable for around 6 months so if you undertake a reasonable amount of skin tests it could be wiser to purchase small volumes of conc extracts which are stable for 2 years and dilute them for use every 3-6 m. If this does not induce a good weal. Continue with each allergen and with the positive and negative controls. Introduce the needle into the skin. Getting started: The fundamentals of why we skin test Intradermal tests may be used to 1) Confirm a clinical dx of atopic dermatitis 2) Enable selection of allergens for immunotherapy Equipment required= 1ml syringes with 25-26g needles. NEVER FROZEN. Sedate the px with a product that has no antihistaminic activity (e. If it is perennial it does not matter. Keep best in glass vials. Generally the px should have been off for at least one week for every month of continual therapy. The most important thing is to ensure that all blebs are the same size. If in doubt give an injection of the positive control (histamine phosphate 1/100. . you will see a rapid blanching of the site. marker pen. e. clippers. without having to shift your hold on the syringe.If atopic dermatitis is still suspected and the problem is seasonal. bevel upwards.05 ml. One advantage is that you can then use the same preparations to make up the vaccine for immunotherapy. Giving the injections: Hold the syringe so that you can inject immediately once the needle enters the skin.If flea allergy is suspected testing with that allergen alone is undertaken. Mark the sites to be injected using a suitable marking pen. xylazine).where more long-acting injectables have been used and possibly less where low dose alternate day therapy has been given.1st rule out other diseases which could account for the clinical signs. Which px should I select for skin testing? . This should cause a readily visible bleb. . Storage of allergens at +4°C. Two common errors: If you are subcutaneous you will not appreciate resistance and there will be no discernible bleb. Clip the hair from the lateral chest. Corticosteroids also interfere with the IDT.000 w/v histamine phosphate. This is the preferred area as it is the most sensitive. good light for reading results. This is the key to successful technique. and inject around 0.g. Preparation of the patient for skin testing: Must have been off antihistamines for 10d. If you inject air. skin testing is best done after the major allergy season.
the punch should be placed in the centre of the lesion. If excisional biopsies are undertaken. that a positive skin test merely indicates that the patient has allergen specific IgE antibodies. The delayed reaction is often very subtle and not obviously urticarial in nature. - . Skin Biopsy Indications for performing a skin biopsy can be summarised as: Any primary eruption of whose nature you are unaware should be sampled. In the case of the flea allergen.Reading the skin test: The time - - Some skin tests are read at 15-20mins after injection Sometimes the immediate reactions are negative. with normal skin excluded (unless the lesion is very small). This is also the case where wedge shaped or elliptical biopsies are concerned. Two approaches to grading Assuming that the size of the histamine weal is +4 and the diluent control is negative. Site selection: Choose primary lesions wherever possible In the case of puch biopsies. reactions can occur either immediately or at 24-48 hours. each reaction is graded on a +1-+4 scale taking into account both the elevation above the skin. with reactions appearing at 4-8 hours. Any dermatosis whose aetiology is still obscure after clinical and other examinations. Alternatively: The diameter of each reaction is measured. Thus if the immediate reaction is negative.g. The results are interpreted having regard to: o The presence of the allergens in the env and o The clinical signs as related to the above. in the case of a suspected tumour normal skin should be included in the margins. the patient should be reassessed at the later time. - Interpreting the test Remember. the degree of erythema and the weal diameter. It does not imply that these are the cause of the disease. Sample a number of lesions in varying stages. The significance of these is unclear and many dermatologists do not routinely record these. e. A positive reaction is any that exceeds 50% of the mean of the weal diameter of the histamine and diluent controls. again.
place on slide. After adherence is assured. loss of hair. it is then transferred to a vial of 10% neutral buffered formalin.Technique: Disinfection of skin is not essential If you chose to do it the skin should be dabbed lightly only with 70% ethanol or with isopropyl alcohol. indicated in majority of cases if not all!!!!! Selection of area to be scraped Choose abnorm skin Select areas where there is excessive scale. superficial first. wedge shaped or elliptical biopsies are taken using a scalpel. 6-8mm disposable punch biopsies are ideal. (A) . No other disinfectants. Do these separately i. response to therapy. then return to do deep. Handle tissue gently.Male scabies mites are on the SURFACE. SURFACE dwellers just visible to the naked eye as grey dots. (D) Superficial and deep scrapings should be taken to cover entire spectrum of parasites. comedones. papules. Same is true of Notoedres. Females burrow into the EPIDERMIS. - . (C) . Dx may not be definitive. moving always the same way until through skin. Skin Scrapings Indications For dx of parasitic dz Cheap and simple.learn from each other Dermopathology is v limited as many dzs look the same.Demodex mites are in the hair follicle. Give pathologist feedback.Cheyletiella on SURFACE and just visible to the naked eye as white dots WALKING DANDRUFF (B) . Working with the pathologist: Form a team and co-operate Choose pathologist with interest in skin dz and who attends derm meetings Give complete hx. your list of ddx’s in rank order. Do not rub! Inject 1-3ml of 2% lidocaine or other suitable LA S/C well clear of the biopsy site. Withdraw punch Pick up biopsy using forceps and handling only the undersurface Excisional. Ear margins are favourites for scabies. lab findings. Location of parasites Harvest mites (“chiggers” or “berry bugs”) are on the SURFACE and just visible as orange-red dots. To prevent distortion (occurs readily with larger biopsies) place biopsy with cut surface down on a piece of cardboard or wood.e.Otodectes usually found in ear but may be on surrounding skin. where the eggs are laid. Gently rotate biopsy punch. .
Conditions amenable to this approach: . If Demodex suspected squeeze skin gently btwn finger and thumb to force any more mites more superficially in the hair follicle Moisten skin with oil (mineral oil.A B C D Technique: Clip hair from area to be scraped If Cheyletiella suspected clip with scissors as otherwise mites will be removed by close use of clippers. corticosteroids. Return for deeper scrapings and continue until there is obvious capillary bleeding. Transfer to slide. If there is a response it is impossible to ascribe the improvement to any one of the drugs administered. Sarcoptes Notoedres Therapeutic trial Specific therapy = dx Combination tx eg abs. and anti-parasitic tx should NOT be used. liquid paraffin) Take a number 10 scalpel blade Blunt the cutting edge on a hard surface Moisten the blade and skin surface with oil and place a drop of oil on the microscope slide. Gently but firmly scrape in the direction of the hair growth at an angle of about 45-70° to the skin surface.
Cheyletiella and harvest mites sometimes hard to confirm. Also sp cautions relating to sheepdogs and other herding breeds. Vit A responsive dermatosis: compare before and after tx Zinc responsive dermatosis: compare before and after zinc sup. In such cases use a combo of suitable parasiticide on pet AND environmental control. . Conditions NOT amenable to this approach: Endocrine dz hypoT. Key concepts Conditions amenable to this approach included: Flea allergy: sometimes evidence of parasite hard to find. Pyodermas: If a bacteria has been isolated from suspected pyoD. Choose approved prod.- - Sarcoptic mange mites often dif to find. Response may be seen in normal animals resulting from supraphysiological doses. Cushings a ddx must be made and if borderline results are obtained rpt testing should be done in 3 months rather than trial tx which could be dangerous. Ivermectin 100% effective but NOT approved for this purpose. Response to tx may be confirmatory. tx with abx to which sensitive (shown in vitro) is best.
This action might not be possible to undo. Are you sure you want to continue?
We've moved you to where you read on your other device.
Get the full title to continue listening from where you left off, or restart the preview.