Lucrare Condur Materiale Si Tehnici ice Engleza

8/3/2019 Lucrare Condur Materiale Si Tehnici ice Engleza

1/5

MATERIALS AND METHODS USED IN CARNIVORESDERMATOLOGIC PATHOPHYSIOLOGY RESEARCH.

PART OF PhD WORK

D. CONDUR, T. PETRU, N. BERCARU, N. VELICU

Facultatea de Medicin Veterinar Spiru Haret

Key words: skin, carnivores, pathology

SUMMARYFor the best therapeutical results, the etiology and the disease mechanism of the involved

disease is needed. The veterinary medic needs a systematic approach, a thorough examination andappropiate diagnostic procedures. The techniques that have been used in this work were: clinicalexamination (including the dermatologic history records), dermatoscopic exam, cytologic exam,

dermatohistopathology, electronic microscopy, serum biochemistry, endocrine tests,hemoleucograms, serum electrophoresis, allergy tests, and other. For the studies, 236 dogs and 87 catswere selected and recorded. All the exams used were useful in etiology and pathophysiology

determination. Not only the skin tests are effective in dermatological diagnostic, but much moreinternal organs disfunctions testing can conduct us to the proper mechanism of disease. Thepathological mechanisms can be multiple, more than one mechanism on the same patient.

The skin is the largest and most visible organ of the body. It is not

only the anatomic and physiologic barrier between animal and environment.

It is synergistic with internal organ systems and reflects pathologic

processes that are either primary elsewhere or shared with other tissues(Scott, 2000).

So we can say the skin is not just an organ with its own reaction

patterns; it also reflects the inside of the body and, in the same time, the

surrounding environment (Gross, 2005).

For the best therapeutical results, the etiology and the disease

mechanism of the involved disease is needed. The veterinary medic needs a

systematic approach, a thorough examination and appropiate diagnostic

procedures (Guaguere, 1999, Hill, 2002).

The techniques that have been used in our work were as follow:

clinical examination (including the dermatologic history records),

dermatoscopic exam, cytologic exam, dermatohistopathology, electronic

microscopy, serum biochemistry, endocrine tests, hemoleucograms, serum

electrophoresis, allergy tests, and other (Wilkerson, 2004).

1


2/5

1. MATERIAL AND METHOD

For our studies, 236 dogs and 87 cats were selected and recorded,

from a total of more than 600 dermatological cases seen. The reason of

selection was the possibilities of the owner to make some clinical exams for

their pet. On the other hand, 37 of the dogs and of the 38 cats with

dermatological problems were homeless animals, catched by us, examed,

threated, and, most of them, given to adoption. The rest remained in our

clinic are waiting to find a careful owner.

On all of our patients clinical examination was done, and the cases

have been recorded on computer, with a case number for each patient.

Where possible, pictures were taken, and lesions maps were completed(fig. 1).

Figure no 1: Skin lesions map Figure no 2: Cytologic microscopical image.

1-keratinocyte with picnotic nuleus; 2-

colagen fiber; 3-Malassezia spp. yeast

The dermatologic history has been taken, except, of course, the

homeless patients. For the last, we were proceeding to a clinical observation

in our clinic.

Dermatoscopic exam has been performed on all cases, also the

cytologic exam. For the dermatoscopy the oil imersion, KaOH 10%, or

lactophenol-blue have been used for sample clarifying.For cytologic exam the Merck Hemacolor and methylene blue

techniques have been used. The Hemacolor coloration has a panoptic result.

The nuclei are coloured red to violet, the cytoplasm pale is blue to reddish,

the keratinocytes blue, the fibers dark blue (fig. 2). Methylene blue has been

used for quick examination samples. The result was dark blue to black

nuclei and pale blue to blue with grannulae cytoplasm.

2


3/5

Dermatohistopathology was done on 135 dogs and 32 cats. Theanesthesia was not usually necesary. The samples were gathered with a

thumb forceps, the specimens have been put in 10% formalin. The methods

used by us were hematoxylin-eosin, tricromic Mallory, toluidine blue, and

anhidrous Giemsa.

Electronic microscopy has been performed on 3 dog cases. The

Babes Institute Electronic Microscopy team helped us in prelevation,

fixation, coloration and interpretation.

Serum biochemistry has been our choice for 200 dogs and 78 cats.

The tests have been performed to the Arkray biochemical device, and to

Synevo human biochemistry laboratories. The most tested parameters were

liver (ALT, AST, bilirubin, total protein, albumin, glucose), and renal (urea,creatinine, phosphorus, calcium).

Endocrine tests were done for 87 dogs and 13 cats suspected to have

hormonal problems. They were performed on Synevo laboratories and on

Innovet Idexx device (for T4 and cortisol dosing). The parameters were

growth hormone (GH), thyroid stimulating hormone (TSH), T3, T4,

Adrenocorticotrop hormone (ACTH), cortisone, testosterone, estradiol, and

progesterone.

Hemoleucograms have been done on all patients. The venous, or

capilar blood was sampled. Panoptical coloration with Hemacolor has been

used.

Serum electrophoresis has been performed on 125 dogs and 24 cats.

Determinations were made using agarose gel electrophoresis machine using

the Tris Barbital buffer at pH 8.6. Serum was diluted 1 / 7 with Tris Barbital

buffer, protein fractions migration was accomplished at 85 V, migration

time beeing 17 minutes and 30 seconds.The coloring of the protein fractions

was performed with Amido Black. After drying, computerisedintegration and calculation of the electroforegrams wasconducted, resulting in 5 distinct protein fractions: albumin,1 globulin, 2 globulin, globulin and globulin.

Allergy tests have been done on 8 dogs and 3 cats. The tests were

made in Innovet veterinary clinic, using an allergy test kit. The hair hadbeen clipped on a rectangular area in a lateral abdominal area, the injection

sites were pointed with a marker, the antigens were injected with insulin

needles. There were a martor (9%0

NaCl solution) and a standard histamin

solution. After two hours, the injected areas were tested for oedema, and the

dimension of tumefactions was measured.

3


4/5

2. RESULTS AND DISCUSSION

All the exams used by us have been useful in etiology and

pathophysiology determination. The clinical examination, together with the

patient history, has been conducting us to next level, laboratory tests, for the

final result of whats happening there. In none of the cases the clinical

examination was performed as single examination.

Dermatoscopic exam was useful in the etiology of 35 cases of dog (9

cats) sarcoptic manges, in 18 cases of canine demodicosis, in 27 cases of

clinical Trychophyton spp. in dogs, 7 cases of cat trychophytosis, 18 casesof Microsporum in dogs, 5 cases of Microsporum in cats.

The cytologic exam helped in 22 cases of clinical Trychophyton in

dogs, 7 cases of cat trychophytosis, 17 cases of Microsporum in dogs, 3

cases of Microsporum in cats, 9 cases of Malassezia infestation in dogs, and

3 in cats. This test helped also in 7 cases of dog parakeratosis, 3 cases of

canine discoid lupus, .

Dermatohistopathology helped us to discover 3 cases of canine

discoid lupus, 7 pemphigus complex in dogs and 3 in cats, 2 in dogs with

bullous pemphigoid, and 1 cat with indolent abcess. It also helped us to see

whats happening in 9 sarcoptic mange infestation in dogs, in 5

demodicoses cases in dogs. in 7 cases of Trychophyton infestations in dogs,5 cases of Microsporum in dog.

Electronic microscopy was helping us for 1 lupus case, and 2

pemphigus complex in dogs.

Serum biochemistry reveled us internal organs functional problems

in 3 lupus affected dogs, in 3 pemphigus complex in dogs, 2 in dogs with

bullous pemphigoid. It also was helpful in 7 cases of hipothyroidism, and in

25 cases of Cushing in dogs, and 1 case of yatrogen hyperprogesteronemia

in 1 tomcat.

Endocrine tests helped us to confirm 7 hipothyroidism cases in dogs,

25 Cushing cases in dogs, hipogonadism in 15 male dogs, 25 female dogs,

10 cases of hipogonadism in female cats.

Hemoleucograms helped us to make an image of white blood cells

evolution in all cases. Neutrophilia was observed in 12 dogs and 4 cats with

immunologic diseases, eosinophilia in 62 external parasitated dogs, in 13

external parasitated cats. Monocytosis was observed in 20 dogs with clinical

micosis. Lymphocytosis was observed in 23 dogs with cronical

4


5/5

dermatologic problems.Serum electrophoresis revealed an increasing of globulin to 1

dog with dermatologic involving of a Walderstrom disease. globulinincreased in all alergic and immunological diseases, with some particular

aspects.

Allergy tests revealed skin allergy in all of 8 dogs an 3 cats tested.

3. CONCLUSIONS

3.1. The ethio-pathogenetic tests on pets are related not only with

investigation possibilities, but with human society, and, why not, with

the economical crysis.3.2. The best solution for a good ethiological and pathogenical

diagnostic is to have all the tests under the same roof, as much as

possible.

3.3. Not only the skin tests are effective in dermatological

diagnostic, but much more internal organs disfunctions testing can

conduct us to the proper mechanism of disease.

3.4. The pathological mechanisms can be multiple, more than

one mechanism on the same patient.

REFERENCES

1. Gross, Thelma Lee, P.J. Ihrke, EmilyJ. Walder,Verena K. Affolter. Skin diseases of the

dog and cat, Second Ed., Blackwell Science Pub., 2005.2. Guaguere E., P. Prelaud A practical guide to feline dermatology english transl., Ed.

Merial, 1999

3. Hill, Peter B.- Small Animal Dermatology: A Practical Guide to Diagnostic Tests, Ed.Butterworth-Heinemann, 2002.

4. Scott Danny W., William H. Miller Jr, Craig E. Griffin - Muller and Kirk's Small AnimalDermatology, VIth Edition, Ed. Saunders, pp. 61-206, 2000.

5. Melinda J. Wilkerson & col. - The immunopathogenesis of flea allergy dermatitis in dogs,an experimental study - Veterinary Immunology and Immunopathology 99, pp 179-192,

2004

5

Lucrare Condur Materiale Si Tehnici ice Engleza

Documents

Transcript of Lucrare Condur Materiale Si Tehnici ice Engleza