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R e n a l Am y l o i d o s i s i n C a t t l e w i t h In a m m a t o r y D i s e a s e s
O.M. Elitok, B. Elitok, and O. Unver
Background: The association of inammatory diseases such as traumatic reticuloperitonitis (TRP), mastitis, metritis, and
pododermatitis with renal amyloidosis in cattle is poorly described.
Hypothesis: Serum amyloid A (SAA) levels are elevated during inammatory diseases, and renal amyloidosis is formed as a
complication.
Animals: This study was conducted with 82 crossbred cattle with mastitis (n 5 18 cows), metritis (n 5 11 cows), TRP (n 5
30 cows), and pododermatitis (n 5 23 : 15 cows and 8 beef cattle). Ten clinically healthy cows served as controls.
Methods: Hematological, urinary, and blood parameters, including SAA, were measured by an automated procedure provided with trade kits. Determination of amyloidal structures was made by histopathological examination of renal biopsy
specimens.
Results: At the end of this trial, amyloidosis was detected in 5 cows displaying typical nephrotic syndrome, with hypoproteinemia and proteinuria in combination with polyuria and weight loss. Furthermore, it was observed that cows with renal
amyloidosis had signicantly higher (P o .01) total leukocyte counts, serum and urine enzyme activities, and urea and creatinine concentrations, with lower serum total protein concentrations, when compared with animals without renal amyloidosis.
Conclusions and Clinical Importance: The incidence of AA amyloidosis in cattle in this study suggests that cattle with
mastitis, metritis, and pododermatitis have a high prevalence of systemic amyloidosis in response to inammation.
Key words: Bovine; Inammation; Renal amyloidosis; Serum amyloid A.
451
Results
aminotransferase (AST), g-glutamyl transferase (GGT), and alkaline phosphatase (ALP) activities were measured by standard
methods with commercial kits supplied by Roche Diagnosticsb in
the Roche/Hitachi 917 Clinical Chemistry Analyzer. SAA concentrations were measured by an enzyme-linked immunosorbent assay
(ELISA) with a Multispecies SAA ELISA kit according to directions supplied by the manufacturer.c
Clinical Variables
Common clinical ndings were anorexia, polyuria
(16%), polydipsia (20%), diarrhea (13%), and weight
loss (100%) (Table 1). The mean baseline temperatures
for all study groups were signicantly higher compared
with the control group (P o .01). Although baseline respiration and heart rate for the 4 study groups and control
group were not different (P 4 .1), animals with mastitis,
TRP, metritis, and pododermatitis had signicantly
decreased rumen motility compared with the control
group (P o .01).
Urinary Examinations
Sterile urine samples were collected via a urinary catheter and
used immediately for analytical tests. Urine samples were analyzed
for GLU, urea, creatinine, and TP concentrations, and AST, GGT,
ALP activities were measured by standard methods with commercial kits supplied from Roche Diagnostics in the Roche/Hitachi 917
Clinical Chemistry Analyzer. Additionally, urinary sediment and
strip testd measurements were evaluated.
Hematological Findings
Histopathology
Statistical Analysis
Statistical analysis was performed using the Statistical Package
for the Social Sciences (SPSS) v. 11.0 for Windows, and data are
listed as the meanSE. For statistical comparisons, Duncan tests
were performed and P values 0.01 were considered statistically
signicant.
Table 1. Clinical variables for the animals with TRP, mastitis, metritis, and pododermatitis and the cattle with renal
amyloidosis.
X SD
Parameters
Temperature (C1)
Pulsation (minutes)
Respiration (minutes)
Rumen contractions (5 minutes)
Subcutaneous edema
Weakness
Inappetence
Polyuria
Polydypsia
Diarrhea
Dehydration
Ascites
Dyspnea
Control,
n 5 10
TRP,
n 5 30
Mastitis,
n 5 18
Metritis,
n 5 11
Pododermatitis,
n 5 23
37.4 0.2b
66 2e
16 2f
9 3a
(
)
(
)
(
)
(
)
(
)
(
)
(
)
(
)
(
)
38.6 0.1a
68 4d
20 4e
5 2b
(
)
(11 ), n 5 30
(11 ), n 5 30
(11 ), n 5 5
(11 ), n 5 6
(1
), n 5 4
(1
), n 5 5
(
)
(1
), n 5 24
38.4 0.4a
71 6bc
23 3c
6 2b
(
)
(11 ), n 5 16
(11 ), n 5 18
(11 ), n 5 5
(11 ), n 5 5
(1
), n 5 4
(1
), n 5 5
(
)
(1
), n 5 6
38.1 0.2a
72 4b
25 2b
5 1b
(
)
(11 ), n 5 11
(11 ), n 5 11
(11 ), n 5 1
(11 ), n 5 3
(1
), n 5 3
(1
), n 5 3
(
)
(1
), n 5 5
38.3 0.1a
70 3c
21 3d
6 2b
(
)
(11 ), n 5 23
(11 ), n 5 23
(11 ), n 5 1
(11 ), n 5 1
(1
), n 5 1
(1
), n 5 2
(
)
(1
), n 5 12
38.4 0.0a
74 4a
26 2a
4 1c
(
)
(11 ), n 5 5
(111), n 5 5
(111), n 5 5
(111), n 5 5
(1
), n 5 5
(11 ), n 5 5
(
)
(1
), n 5 5
612
510
2446
1012
2040
4060
410
02
01
Reference Range
7.2 1.0b
7.3 1.2e
32.2 2.4b
8.2 0.4a
32 3a
60 2c
6 3a
2 1a
00
Control, n 5 10
4.8 1.2d
9.6 1.4c
30.4 3.2c
6.6 0.2b
20 4d
78 8a
2 0b
0 0b
00
TRP, n 5 30
4.4 1.1e
10.2 1.2b
30.6 2.0c
6.6 0.6b
28 2b
72 1b
0 0c
0 0b
00
Mastitis, n 5 18
5.2 1.0c
9.1 1.6b
30.8 2.6c
6.8 0.2b
26 2bc
73 2b
1 0c
0 0b
00
Metritis, n 5 11
5.1 1.4c
9.0 1.0d
30.2 1.8c
6.8 0.0b
24 4c
73 1b
3 0b
0 0b
00
Pododermatitis, n 5 23
8.0 1.2a
14.3 1.1a
48.2 1.8a
14.2 0.2b
20 3c
78 1b
1 0c
0 0b
00
88.40176.8
6775
3036
2030
78132
1319
0488
Variable
2.57.5
Creatinine (mmol/L)
TP (g/L)
AB (g/L)
Urea (mmol/L)
AST (U/L)
GGT (U/L)
ALP (U/L)
SAA (mg/mL)
Glucose (mmol/L)
96.4 15.6
70 1.6a
32.3 1.4a
8.5 0.8e
88.2 12.2d
17.1 0.4d
236.6 22.3e
17.2 1.3c
3.1 0.4a
Control, n 5 10
368.41 16.1
58.3 2.4c
22.4 1.3d
16.8 1.4b
184.3 21.6b
43.3 5.5b
681.3 34.6b
68.4 1.6b
1.9 0.2b
TRP, n 5 30
382.41 12.3
54.1 1.6d
19.4 1.6e
14.6.4 1.1c
188.4 18.5b
44.1 3.4b
677.7 31.2b
68.3 1.7b
2.0 0.0b
Mastitis, n 5 18
353.28 21.4
63.4 3.8b
25.3 1.8c
11.2 1.8d
156.2 14.8c
38.1 3.6c
644.6 45.2c
66.4 2.0b
2.0 0.0b
Metritis, n 5 11
355.41 18.2
64.2 1.1b
29.3 2.1b
11.4 1.3d
152.8 17.2c
36.2 2.4c
629.4 28.6d
66.2 1.8b
1.9 0.3b
Pododermatitis, n 5 23
856.33 48.4a
34.3 1.8e
18.9 0.7f
41.4 2.7a
356.4 35.8a
104.3 4.6a
985.6 64.3a
79.4 2.0a
1.6 0.2c
Reference Range
Parameters
X SD
Table 3. Metabolic prole for the animals with TRP, mastitis, metritis, and pododermatitis and the cattle with renal amyloidosis.
Erythrocyte (106/mm3)
Total leukocyte (103/mm3)
PVC (%)
Hemoglobin (g/dL)
% Neutrophil
%Lymphocyte
%Eosinophil
%Monocyte
%Basophil
Parameters
X SD
Table 2. Hematological ndings for the animals with TRP, mastitis, metritis, and pododermatitis and the cattle with renal amyloidosis.
452
Elitok et al
453
Table 4. Urinary variables for the animals with TRP, mastitis, metritis, and pododermatitis and the cattle with renal
amyloidosis.
X SD
Control,
n 5 10
Parameters
Polyuria
Granular cast
Crystals
Pyuria
TP (g/L)
Urea (mmol/L)
GGT (U/L)
ALP (U/L)
AST (U/L)
Creatinine (mmol/L)
Specic gravity
(
(
(
)
)
)
0.0d
2.1 0.3e
0.4 0.01e
0.1 0.04f
0.4 0.01d
0.0e
1.030b
TRP,
n 5 30
Mastitis,
n 5 18
Metritis,
n 5 11
Pododermatitis,
n 5 23
(1
)
(1
)
(1
)
(1
)
16.9 1.bc
6.6 0.9b
18.6 1.3c
136.4 12.6b
36.4 1.3b
44.1 4.3c
1.010d
(1
)
(1
)
(1
)
(1
)
17.6 1.3b
6.2 0.5c
21.4 2.0b
126.3 11.0c
38.3 2.0b
48.4 5.2b
1.010d
(1
)
(1
)
(1
)
(1
)
15.4 1.2c
6.0 0.6cd
13.1 1.2d
110.0 14.1d
28.0 1.1c
36.6 6.0d
1.020c
(1
)
(1
)
(1
)
(1
)
14.8 1.3c
5.8 0.5d
14.1 1.1d
100.8 14.6e
26.3 0.8c
37.1 3.0d
1.020c
(11 )
(11 )
(11 )
(111)
28.3 2.2a
14.8 1.5a
56.4 1.8a
323.4 32.1a
264.8 44.6a
87.8 6.7a
1.050a
Urinary Findings
Proteinuria and lower urine specic gravity values
were the most prominent signs of urinalysis in this trial.
Additionally, higher urinary urea and creatinine concentrations along with higher enzyme activities for GGT,
AST, and ALP were determined as the other conspicuous
signs. In the dipstick examination of urine samples, proteinuria (1
), pyuria (1
), and casts were
determined for the study groups (Table 4).
Histopathological Findings
All the animals survived for 1 week after performing
percutaneous kidney biopsy, and none of them showed
untoward adverse effects during the study. Two cows
from the TRP group and one from the pododermatitis
group were shown to have renal amyloidosis. Lesions
were most notable in the glomeruli (Table 2), where amyloid was present in capillary walls and mesangium
in 5 of 82 animals (6%). A periglomerular mononuclear
cell interstitial inltrate was occasionally noted around
the most severely damaged glomeruli, and inammatory
cells consisted predominantly of lymphocytes and plasma cells (Fig 1). Although amyloid deposits were
hypocellular, a periglomerular mononuclear cell interstitial inltrate was occasionally noted around the most
severely damaged glomeruli. Other lesions detected
in histopathology in the present study are listed in
Table 5.
454
Elitok et al
Table 5. Histopathological ndings in animals with TRP, mastitis, metritis, and pododermatitis and the cattle with
renal amyloidosis.
Control,
n 5 10
Parameters
Glomerulo-tubulonephrosis
Tubuler degeneration and necrose
Glomerular atrophy and degeneration
Chronic glomerulonephritis
Thickening of basal membrane
Periglomerular lymphohystiyociter inltration
Desquamation of tubulus epithel
Amyloidosis
Glomerular amyloid degeneration
TRP,
n 5 30
Mastitis,
n 5 18
Metritis,
n 5 11
Pododermatitis,
n 5 23
18 (111)
12 (111)
9 (11)
8 (11)
2 (11)
15 (11)
3 (11)
2
2 (111)
4 (111)
4 (111)
14 (11)
10 (111)
10 (1)
10 (111)
4 (111)
2
2 (111)
5 (11)
3 (1)
1 (1)
1 (1)
3 (1)
6 (1)
1 (11)
( )
( )
13 (111)
8 (11)
7 (1)
3 (11)
4 (11)
8 (111)
2 (11)
1
1 (111)
5 (111)
5 (111)
5 (111)
5 (111)
5 (111)
5 (111)
5 (111)
5 (111)
5 (111)
Arabic numerals show the number of animals and marks in the parentheses emphasize severity of the lesions: ( ), absent; (1), mild; (11),
moderate; (111), severe.
The amount of amyloid in the kidney as detected by Congo red stain was rated as 1, mild deposition; 11, moderate deposition; 111,
extreme deposition. Furthermore, other lesions detected in histopathology in the present study are scored according to severity of the lesions
as 1, mild; 11, moderate; 111, severe.
TRP, traumatic reticuloperitonitis.
Discussion
This study was undertaken to obtain epidemiological
information about secondary amyloidosis in Turkey,
with a special focus on etiological distribution. This is a
comprehensive pathologic study of AA amyloidosis in
cattle with mastitis, TRP, metritis, and pododermatitis.
Amyloid deposits were found in glomeruli by microscopic examination of the kidneys, and with Congo red
staining renal amyloidosis was conrmed, as previously
reported incidentally in cows.1620 It has been reported
that only in the most advanced cases, with the glomerulus
being in a terminal stage, was a mononuclear cell interstitial inltrate noted.6,7 In this study, we found that the
amyloid deposits were located only within the glomeruli
and that inammatory cells consisted predominantly of
lymphocytes and plasma cells, as has been previously
found in cattle and in dogs.16,18,21
AA amyloidosis characteristically develops in humans
and animals as a consequence of long-standing, chronic
infectious or inammatory processes. It has also been
described in a few cases of bovine spongiform encephalopathy.2224 In the present study, the animals suffered
from mastitis, TRP, metritis, and pododermatitis that
had been caused predominantly by bacterial agents. Bacterial origins have previously been reported in cases of
chronic suppurative pneumonias of domestic and wild
small ruminants.17,20,25,26 However, both medical histories and the laboratory results in this study indicated that
all diseases were in a chronic stage.
In this study, it was determined that SAA levels in
mastitis and TRP groups were higher than in the other
groups. Similar ndings for mastitis were previously reported by Gronlund et al.27 Acute-phase proteins such as
SAA have been identied as markers of inammation in
cattle because they are produced by the liver in response
to proinammatory cytokines.8,28 Additionally, numerous reports on the use of acute-phase protein responses
to predict severity or chronicity of cattle diseases
have been published.2932 Viral infections generally lead
Footnotes
a
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