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Marcin Jankowski, Agnieszka Halon*, Krzysztof Kubiak, Jolanta Spuzak, Jozef Nicpon, Igor Maksymovych**, Urszula Paslawska, Kamila Glinska-Suchocka ®
Department of Internal and Parasitic Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life
Sciences, pl. Grunwaldzki 47, 50-366 Wroclaw, Poland *Department of Pathological Anatomy, Medical University, ul. Marcinkowskiego 1,
50-368 Wroclaw, Poland **Department of Clinical Diagnostics, Lviv National University of Veterinary Medicine and Biotechnology, ul. Pekarska 50, 79010 Lviv, Ukraine
DIAGNOSIS OF GLOMERULONEPHRITIS IN DOGS
Key words: glomerulonephritis, dog
Introduction
Glomerulonephritis is a disease which leads to disorders in the normal structure of glomeruli manifested by a decrease in renal filtration. At present it is considered to be the most common cause of chronic renal insufficiency in dogs. This condition is most frequently related to the presence of immunological complexes in the wall of glomerulus capillaries which promote a complex inflammatory reaction. Complement, neutrophils, macrophages, platelets, mesangial cells and endothelial cells of the glomerulus vessel are involved in this reaction. As a result, the thickening of capillaries' walls, multiplication of mesangial cells or both phenomena may by take place (5, 9, 12, 17, 18, 19, 20).
Glomerulonephritis is a polyetiological disease caused by: bacterial infections most frequently resulting from such diseases as: endocarditis, brucellosis, pyometra, boreliosis, mycoplasmal polyarthritis, prostatitis, sepsis; viral infections, eg.: Rubarth's disease; parasitic diseases, eg.: dirophilariosis, trypanosomatosis, leishmaniosis; all kind of tumors, pancreas diseases, chronic skin diseases, Cushing's syndrome, diabetes, hypothyroidism (1, 2, 5, 6, 10,15, 22, 23).
Considering etiology of this disease, its course and complex pathogenesis, it should be stated that intravital diagnosis of glomerulonephritis in dogs is relatively difficult.
Task, the aim of the article
The aim of the undertaken study was to determine the possibility of intravital diagnosis of glomerulonephritis in dogs.
Material and methods
The study was conducted on 15 dogs of different breed and sex, aged 11 months to 11 years, in which glomerulonephritis had been suspected and which had been referred to Department of Internal and Parasitic Diseases with Clinic for Horses, Dogs and Cats.
® Marcin Jankowski, Agnieszka Halon*, Krzysztof Kubiak, Jolanta Spuzak, Jozef Nicpon, Igor Maksymovych**, Urszula Paslawska, Kamila Glinska-Suchocka, 2008
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All the animals were subject to anamnesis and clinical examination, as well as additional examinations, such as: blood laboratory tests (hematological and biochemical), urinalysis, renal ultrasound examination, kidney biopsy and histopathological evaluation of bioptates. In hematological tests the following parameters were determined: erythrocytes count (RBC), leucocytes count (WBC), hemoglobin concentration (HGB), hematocrit value (Hct), red blood cells indices (MCV, MHC, MCHC), platelets count (PLT), coagulation time and bleeding time. In biochemical tests the following parameters were determined: activity of: aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), concentration of: urea, creatinine, total protein, albumins, glucose, cholesterol, calcium (Ca++), inorganic phosphorus (Pn), natrium (Na+), kalium (K+), and chlorine (Cl-). In the physico-chemical examination of the urine the attention was paid to colour, specific weight, pH, the presence of: protein, glucose, acetone, blood, urobilinogen and bile pigments, and in the urine sediment - to the presence of: erythrocytes, leucocytes, epithelial cells, mineral components and bacteria.
The ultrasound examination of kidneys was performed with USG Digi prince DP-3300 Vet apparatus with a 5 MHz miniconvex head. The location of kidneys, their size, shape and internal structure were evaluated in this examination.
Kidney bioptates were collected using semi-automatic Tru-cut needles of the following parameters: diameter 1.6 mm, lenght - 150 mm, the chamber lenght for the tissue sample - 20 mm.
The collected bioptates were fixed in 10% buffered formalin solution, and embedded in paraffin blocks. This paraffin blocks were cut into 5 ^m thick slices using a microtome. Next, paraffin was removed from samples in xylene and samples were placed on Super Frost slides. The staining with hematoxylin and eosin, p.a.S. method, Masson method and Jones method was performed according to the histopathological laboratory protocol.
The bioptates containing minimum 5 glomeruli were considered as representative (13). The pathological changes of glomeruli were evaluated basing on the classification of primary and secondary glomerulopathies according to McGavin et al. (16) and Janette et al. (11) .
Results of resarch
In anamnesis and clinical examination in dogs the following clinical signs were observed: decreased appetite, anorexia, polydypsia, polyuria, pollakiuria, decreased mobility, weight loss, opisthotonus, posterior ataxia, vomiting, tenderness in the kidney area, ascites and congestive edema. It should be noted that clinical signs in these dogs were non-specific and were related to mane systems and organs. Similar clinical symptoms in dogs with glomerulonephritis were reported by other authors (4, 21, 24). Tab.1. present percentage part of individual clinical symptoms in dogs with suspected glomerulonephritis.
In the hematological examination a decreased erythrocytes count and decreased hematocrit value was observed in 5 (33.3%) dogs. A decreased hemoglobin concentration was observed in 3 (20.0%) dogs. Two (13.3%) dogs demonstrated decreased MCH i MCHC and 4 (26.6%) dogs - increased leucocytes count. The
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remaining hematological parameters were within the physiological norm. It should be noted that only 33.3% of dogs showed sings of non-regenerative anemia accompanying chronic renal insufficiency, the cause of which may be glomerulonephritis.
The biochemical blood tests showed increased values of the following parameters: urea - 9 (60.0%) cases, creatinine - 10 (66.6%) cases, non-organic phosphorus - 4 (26.6%) cases and a decrease in total protein value - 5 (33.3%) cases and albumins - 5 (33.3%) cases. The remaining blood biochemical parameters were within the physiological norm. It should be emphasized that a decrease in the total protein and albumins concentration considered as typical of glomerulonephritis also occurred only in 33.3% of dogs.
The physico-chemical urine examination showed: turbid urine - 5 (33.3%) cases, slightly turbid urine - 1 (6.6%) case, decreased urine specific weight - 7 (46.6%) cases, change of urine pH - 7 (46.6%) cases, proteinuria - 13 (86.6%) cases, presence of blood - 5 (33.3%) cases. In the urine sediment fresh erythrocytes were observed in 6 (40.0%) dogs, lixiviated erythrocytes - in 4 (26.6%) dogs, leucocytes -in 4 (26.6%) dogs, transitional epithelium - in 2 (13.3%) dogs, round epithelium - in 5 (33.3%) dogs, ammonium-magnesium triphosphates crystals - in 2 (13.3%) dogs, calcium oxalate crystals - in 3 (20.0%) dogs, urine casts - in 2 (13.3%) dogs and bacteria - in 4 (26.6%) dogs. It was found that in dogs proteinuria is the most common sign in urinalysis indicating glomerulonephritis, which is confirmed by observations of numerous authors (3, 7, 8, 14).
The ultrasound examination revealed disorders in the normal kidney picture in 8 (53.0%) cases; these were: enlargement of one or two kidneys - 2 cases, an increase in renal parenchyma echogenicity - 2 cases, blurred renal parenchyma structure - 4 cases, thickening of renal cortex - 2 cases, thickening of renal capsule - 1 case and fibrosis of renal limiting membrane - 3 cases.
All the collected bioptates fulfilled diagnostic criteria and contained renal cortex, single arteries and renal medulla. The lenght of the collected kidney bioptates ranged from 5 mm to 20 mm (mean: 14 mm). The bioptates included 4 - 24 glomeruli in the renal cortex part (mean: 11 glomeruli). Only 1 (6.6%) bioptate contained fewer than 5 glomeruli. Zatelli et al. (25) obtained a similar percentage (3.9%) of bioptates with fewer than 5 glomeruli in their studies.
Out of 15 performed oligobiopsies the significant pathological changes of glomeruli were observed in 13 (86.6%) bioptates. Six following histopathological diagnoses were made: focal segmental glomerulosclerosis (glomerulosclerosis focalis et segmentalis) - 3 (23.1%) cases, membranoproliferative glomerulonephritis (glomerulonephritis mesangio-capillaris) - 4 (30.7%) cases, glomerulonephritis mesangialis (glomerulonephritisploriferativa mesangialis) - 2 (15.4%) cases, chronic diffuse glomerulonephritis (glomerulonephritis chronica diffusa) - 2 (15.4%) cases, extracapillary glomerulonephritis (glomerulonephritis proliferativa extracapillaris) -1 (7.7%) case, minimal change disease (glomerulonephritis submicroscopica) - 1 (7.7%) case.
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Only in 2 (13.3%) cases the histopathological examination of bioptates did not show any pathological changes in the kidney tissue visible in the light microscope.
The presence of the glomerulopathy interstitial component in the form of dispersed and/or focal inflammatory infiltration consisting predominantly of lymphocytes with additive of plasmatic cells was found in 5 (38.4%) cases. The features of arterial vessels walls hyalization of different degree were also found in 5 (38.4%) cases. Focal or diffused stroma fibrosis of small or moderate intensity was observed only in 3 (23.1%) cases. In 9 (69.2%) examined kidney bioptates the presence of morphological exponents of moderate or severe proteinuria in the form of protein degeneration of tubules epithelium (athrocytosis) and intratubular protein casts were observed. In 1 case the foci of toxic damage of renal tubular epithelium in the form of acute tubular necrosis were observed in the renal interstitial tissue.
Conclusions
The conducted studies led to the conclusion that the clinical signs accompanying glomerulonephritis are non-specific and may indicate the diseases of many systems and organs. Moreover, it was found that the most specific laboratory parameter indicating glomerulonephritis in dogs is the presence of protein in urine (proteinuria) and the best intravital diagnostic method enabling final diagnosis of glomerulonephritis in dogs is oligobiopsy and histopathological evaluation of bioptates.
Reference
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2. Aresu L., Valenza F., Ferroglio E., Pregel P., Uslenghi F., Tarducci A., Zanatta R.: Membranoproliferative glomerulonephritis type III in a simultaneous infection of Leishmania infantum and Dirofilaria immitis in dog. J. Vet. Diagn. Invest. 2007, 19, 569 - 572.
3. Biewenga W. J., Gruys E.: Proteinuria in dog: a clinicopathological study in 51 proteinuric dogs. Res. Vet. Sic. 1986, 41, 257 - 264.
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22. Sanchez-Cordon P. J., Salguero F. J., Nunez A., Gomez-Villamandos J. C., Carrasco L.: Glomerulonephritis associated with simultaneous canine adenovirus-1 and Dirofilaria immitis infection in a dog. J. Vet. Med. 2002, 49, 235 - 239.
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25. Zatelli A., Bonfanti U., Santilli R., Borgarelli M., Bussadori C.: Echo-assisted percutaneous renal biopsy in dog. A retrospective study of 229 cases. Vet. J. 2003, 166, 257 - 264.
Tab. 1. Percentage part of individual clinical symptoms in dogs with suspected glomerulonephritis.
□ zmniejszony apetyt
□ brak apetytu
□ wzmozone pragnienie
□ wielomocz
□ cz^stomocz
□ spadek aktywnosci ruchowej
□ spadek masy ciala
□ wygi^cie grzbietu do gory ■ niezbornosc ruchowa
□ wymioty
□ bolesnosc okolicy nerek
□ wodobrzusze
□ obrz^ki zastoinowe
Summary
The aim of the undertaken study was to determine the possibility of intravital diagnosis of glomerulonephritis in dogs. The study was conducted on 15 dogs of different breed and sex, aged 11 months to 11 years, in which glomerulonephritis had been suspected. All the animals were subject to anamnesis and clinical examination, as well as additional examinations, such as: blood laboratory tests (hematological and biochemical), urinalysis, renal ultrasound examination, kidney biopsy and histopathological evaluation of bioptates.
The conducted studies led to the conclusion that the clinical signs accompanying glomerulonephritis are non-specific and may indicate the diseases of many systems and organs. Moreover, it was found that the most specific laboratory parameter indicating glomerulonephritis in dogs is the presence of protein in urine (proteinuria) and the best intravital diagnostic method enabling final diagnosis of glomerulonephritis in dogs is oligobiopsy and histopathological evaluation of bioptates.
Стаття надшшла до редакцИ 20.09.2008
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