PROCEDURES PRO h DIAGNOSTICS/DERMATOLOGY h PEER REVIEWED

Trichograms
Katrine Voie, DVM, MS, DACVD
Gulf Coast Veterinary Specialists
Houston, Texas

dF
 IGURE 1 Adult Demodex canis mite seen
on a trichogram

A trichogram is the evaluation of hairs using a micro- pected demodicosis, approximately 30 to 40 hairs per
scope and can be a valuable diagnostic tool in any derma- affected body location should be collected and evaluated;
tology case. A trichogram can help the practitioner however, collecting at least 30 to 40 hairs in all canine or
examine hairs for parasites (eg, Demodex spp [Figure 1]) feline cases can help ensure a representative sample.
that may be adjacent to the root of the hair or attached to
the hair itself (eg, louse nits [Mallophaga, Anoplura]); In cases of suspected demodicosis, the author prefers to
changes in the cortex of the hair, as can occur with der- perform deep skin scrapings over most of the body
matophytosis; fractures in the hair tips due to alopecia; rather than to perform a trichogram; however, in the
pigmentary changes; and other conditions and/or abnor- author’s experience, trichograms are preferably con-
malities.1 The hair roots can also be examined to deter- ducted in all cases around the eyes, where scalpel blades
mine if the hairs are in anagen (ie, round, smooth, often in a nonsedated patient can be dangerous, or over bony
pigmented bulbs) or telogen (ie, club- or spear-shaped prominences, where there is not a flat area that allows
bulbs) and if the patient is barbering its coat, as barbered for skin scraping. The nail beds are a good location in
hairs appear shorter and split or fractured due to which to perform a trichogram because the digits are
trauma.1,2 common areas for demodectic mange in dogs or derma-
tophytosis in cats1,3 yet are difficult areas in which to
If viable, hairs should be plucked from active lesions (eg, perform skin scrapings. When performing a trichogram
lesions of alopecia, crusting, scaling, or follicular cast- on the nail beds, plucking hairs from multiple toes can
ing), and plucking vibrissae (ie, whiskers) should be ensure that representative samples from any affected
avoided. If patchy alopecia is present, collecting hairs areas are obtained.
from the margin of an alopecic lesion is best. However,
central hair regrowth may be present on the lesion in The hairs should then be placed on a prepared glass
areas of healing, in which case collecting from this area slide. For most preparations, mineral oil is used to evalu-
may not aid in diagnosis. Collecting hairs from the lead- ate trichograms, although a 10% to 20% potassium
ing edge of the lesion (similar to collecting samples for hydroxide solution may be used to evaluate hairs for
cytologic evaluation) may allow for a better representa- arthrospores in cases of suspected dermatophytosis.3
tion of the active process of the lesion. In cases of sus- The author suggests placing the roots of the hairs in the

18 cliniciansbrief.com June 2017

center of the mineral oil if evaluating for Demodex spp. melanin can be seen distributed unevenly
The hairs, if being evaluated for dermatophytosis or throughout the hairs.7 Skin biopsy is recom-
structural abnormalities, should be oriented in a length- mended in these cases for definitive diagnosis.
wise fashion (ie, parallel to the long end of the slide) so
they do not fall over the edge of the slide. For cases of
suspected demodicosis, low power (10×), low light, and a A
lowered condenser will allow for better visualization.4

If the light is too high, it may illuminate through the

bodies of the Demodex spp mites and make them difficult
to visualize, whereas lower light allows the light to
refract off them (Figure 2). Demodex spp mites—which
should be categorized as adults, nymphs, larvae, or eggs
and as either dead or alive—should be counted in the
same way as with skin scrapings.

Demodicosis can be diagnosed via trichogram in many

cases. As compared with deep skin scrapings, tricho-
B
grams have an approximately 75% to 80% sensitivity for
diagnosing Demodex spp mites.5 The mites that live in
the hair follicles become dislodged when the hairs are
plucked for a trichogram and settle onto the slide next to
the hair root when placed in mineral oil.

Dermatophytosis can be diagnosed via fungal culture6;

however, Wood’s lamp examinations can be useful in
certain cases, as hairs affected with certain dermato-
phyte species produce a metabolite that causes the hairs d FIGURE 2 (A) Trichogram (10×) showing Demodex spp
to glow when exposed to the light.6 Trichograms can be mites from a dog. Note how light affects the visualiza-
useful in cases in which ringworm is suspected, as cer- tion of mites. The high light and high condenser create
poor visibility. (B) The same field of the trichogram pic-
tain changes visible to a trained eye via trichogram may tured in Figure A but with low light and a lower con-
assist in making a more rapid diagnosis. Hair can be denser, which allow for excellent visualization
plucked from areas of alopecia, scaling, or crusting. The
author typically evaluates the hairs first under 10 times
magnification for any abnormalities and then under 40
times magnification to better evaluate the hairs. Hairs
should be evaluated for a classic soggy log appearance (ie,
fractured, swollen, flaky-appearing cortex) where ecto-
thrix arthrospores are seen along the outer edge of the
hair or where spores or mycelia are seen within the hair.6

In cases of follicular dysplasia (ie, color-dilution alopecia

or black-hair follicular dysplasia), melanin clumping can
be evident during hair examination (Figure 3). Melanin
dF
 IGURE 3 Microscopic image of a hair from a dog with
pigment is usually distributed fairly evenly throughout color-dilution alopecia. Large clumps of melanin in the
the length of the hair. In these cases, large clumps of hair shaft can be seen.

June 2017 cliniciansbrief.com 19

PROCEDURES PRO h DIAGNOSTICS/DERMATOLOGY h PEER REVIEWED

STEP-BY-STEP
WHAT YOU WILL NEED
TRICHOGRAM
h Hemostats (preferably curved
hemostats, as they are easier
to hold parallel to the skin
surface, thereby allowing STEP 1
removal of more hairs at once)
h Glass slides After performing a dermatologic examination, identify the affected areas
and determine whether a trichogram will be helpful to the case (ie, if
h Mineral oil or potassium
demodicosis, dermatophytosis, or lice is suspected; if patient is affected in
hydroxide
difficult locations [eg, around the eyes, toes]). Then, using curved hemo-
h Cover slips
stats, pluck approximately 30 to 40 hairs from each active lesion (eg, lesions
h Microscope of alopecia, crusting, scaling, or follicular casting) and/or body location.

STEP 2
Place the hairs on a glass slide with either mineral oil or 10% to 20%
potassium hydroxide solution. Cover with a cover slip.

Author Insight
References If examining for demodicosis in long-haired patients, it may
1. Scott DW, Miller WH, Griffin CE.
Diagnostic methods. In: Scott DW, Miller
be helpful to trim the ends of the hairs with scissors before
WH, Griffin CE, eds. Muller & Kirk’s Small
Animal Dermatology. 6th ed. St. Louis,
plucking to avoid having large clumps of hair hanging over
MO: Eslevier Saunders; 2001:87-91, 231-
239, 309-310.
the edge of the slide.
2. Diaz SF, Torres SM, Dunstan RW, Jessen
CR. The effect of body region on the
canine hair cycle as defined by unit area
trichogram. Vet Drmatol. 2004;15(4):225- STEP 3
229.
3. Frymus T, Gruffydd-Jones T, Pennisi Examine hairs under a microscope. Evaluate:
MG, et al. Dermatophytosis in cats:
ABCD guidelines on prevention and h Spaces between the hairs for any evidence of parasites
management. J Feline Med Surg.
h Hair bulbs to determine anagen or telogen phases, if desired
2013;15(7):598-604.
4. Hnilica KA, Patterson AP. Diagnostic h Hair shafts for any abnormalities in the cortex or excessive clumping of
techniques. In: Hnilica KA, Patterson AP,
eds. Small Animal Dermatology: A Color
melanin
Atlas and Therapeutic Guide. 4th ed. St.
Louis, MO: Elsevier; 2017:30-31.
5. Saridomichelakis MN, Koutinas AF, Author Insight
Farmaki R, Leontides LS, Kasabalis D.
Relative sensitivity of hair pluckings and
exudate microscopy for the diagnosis
Too much light from a microscope may affect the practitioner’s
of canine demodicosis. Vet Dermatol.
2007;18(2):138-141.
ability to visualize the hairs and any abnormalities (Figure 2,
6. Moriello KA, DeBower DJ. Cutaneous previous page). n
fungal infections. In: Greene CE, ed.
Infectious Diseases of the Dog and Cat.
4th ed. St. Louis, MO: Eslevier; 2012:588-
602.
7. Laffort-Dassot C, Beco L, Carlotti DN.
Follicular dysplasia in five Weimaraners.
Vet Dermatol. 2002;13(5):253-260.

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