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Histopathological, immunohistochemical, and parasitological studies on pathogenesis of Coenurus cerebralis in sheep

Yilmaz Rahsan
Yilmaz Rahsan
, 
Yumusak Nihat
Yumusak Nihat
, 
Yilmaz Bestami
Yilmaz Bestami
, 
Ayan Adnan
Ayan Adnan
 and 
Aysul Nuran
Aysul Nuran
   | Mar 30, 2018
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Published Online: Mar 30, 2018
Page range: 35 - 41
Received: Dec 19, 2017
Accepted: Mar 09, 2018
DOI: https://doi.org/10.2478/jvetres-2018-0005
Keywords
© 2018 Y. Rahsan et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

A – macroscopic appearance of cysts in the brain and cerebellum. Coenurus cysts in the right frontal lobe (black arrow) and in the left cerebellum (white arrow). B – microscopical appearance of the inflamed area around the cyst in the brain. Calcification (thin arrow), mononuclear cell infiltration (white arrow), hyperaemia (arrowhead), H&E, 100×. C – giant cell (thin arrow), mononuclear cell infiltration (asterisk), necrosis (white arrow), hyperaemia (arrowhead), H&E, 100×
A – macroscopic appearance of cysts in the brain and cerebellum. Coenurus cysts in the right frontal lobe (black arrow) and in the left cerebellum (white arrow). B – microscopical appearance of the inflamed area around the cyst in the brain. Calcification (thin arrow), mononuclear cell infiltration (white arrow), hyperaemia (arrowhead), H&E, 100×. C – giant cell (thin arrow), mononuclear cell infiltration (asterisk), necrosis (white arrow), hyperaemia (arrowhead), H&E, 100×

Fig. 2

2% gel electrophoresis image of some positive samples in the PCR process (446 bp)
2% gel electrophoresis image of some positive samples in the PCR process (446 bp)

Fig. 3

Microscopical appearance of positive staining cells CD3, CD79α, CD163, and GFAP. A – CD3 positive cells (asterisk), necrosis (black arrow), IHC staining, 100×. B – CD3 positive cells (black arrow), IHC staining, 600×. C – CD79α positive cells (black arrow), IHC staining, ×400. D – CD79α positive cells (black arrow), IHC staining, 600×. E – CD163 positive cells (black arrow), IHC staining, 400×. F – CD163 positive cells (black arrow), IHC staining, 600×. G – GFAP positive cells (black arrow), mononuclear cell infiltration (asterisk) IHC staining, 200×. H – GFAP positive cells (black arrow), IHC staining, 600×
Microscopical appearance of positive staining cells CD3, CD79α, CD163, and GFAP. A – CD3 positive cells (asterisk), necrosis (black arrow), IHC staining, 100×. B – CD3 positive cells (black arrow), IHC staining, 600×. C – CD79α positive cells (black arrow), IHC staining, ×400. D – CD79α positive cells (black arrow), IHC staining, 600×. E – CD163 positive cells (black arrow), IHC staining, 400×. F – CD163 positive cells (black arrow), IHC staining, 600×. G – GFAP positive cells (black arrow), mononuclear cell infiltration (asterisk) IHC staining, 200×. H – GFAP positive cells (black arrow), IHC staining, 600×

Difference between positive stained cell numbers using different antibodies

Antibody X ±SE Min Max
CD3 31.68 ± 2.362a 22.6 51.2
CD79 18.627 ± 0.705b 13.4 24.2
CD163 57.147 ± 3.68c 30.4 80.0
GFAP 85.347 ± 0.609d 80.0 89.0
P < 0.001

Correlation between positive stained cell numbers using different antibodies

Antibody CD3 CD79 CD163
CD3
CD79 0.351
CD163 –0.218 –0.059
GFAP 0.0214 –0.024 0.14

Cyst location and scolex numbers

No Sex Age Cyst location Size of cyst Scolex number
1 Male 1 year Left parietal lobe 6×5 cm 53
Right parietal lobe 5×4 cm 47
2 Female 6 months Left parietal lobe 4×3 cm 38
Right parietal lobe 4×3 cm 36
3 Male 9 months Left frontal lobe 5×4.5 cm 49
Right frontal lobe 3×2 cm 26
Right parietal lobe 3×2 cm 29
4 Female 2 years Left parietal lobe 6×5 cm 55
Right parietal lobe 5×4 cm 45
5 Male 6 months Right parietal lobe 3×2.5 cm 32
Right temporal lobe 3×2 cm 24
Left frontal lobe 3×2 cm 25
Left parietal lobe 3×2 cm 28
6 Male 1 year Left frontal lobe 5×4.5 cm 38
Right frontal lobe 5×5 cm 48
7 Female 1 year Left frontal lobe 6×5 cm 53
Right frontal lobe 4×5 cm 50
8 Female 4 years Left frontal lobe 4×3 32
Right frontal lobe 4×3 30
9 Male 6 months Left frontal lobe 3×3 28
Right frontal lobe 3×3 26
Left parietal lobe 2×2.5 21
Right parietal lobe 2×2.5 24
10 Male 8 months Left temporal lobe 4×3 36
11 Male 6 months Left temporal lobe 3×3 33
Right temporal lobe 3×2 21
12 Female 2.5 years Left occipital lobe 3×3 33
Right occipital lobe 3×2 25
13 Male 7 months Right frontal lobe 3×3.5 cm 38
Cerebellum 2×2 cm 30
14 Male 7 months Cerebellum 3×2 cm 30
15 Female 1 year Left frontal lobe 5×6 cm 47
Right frontal lobe 4×3 cm 35

Differences in immunohistochemical staining protocol

CD3 CD79α CD163 GFAP
Antigen retrieval 20 min, 120°C, 1 atm, autoclave, citrate buffer 20 min, 120°C, 1 atm, autoclave, citrate buffer 10 min, 800 watt, microwave, citrate buffer 15 min, room temperature, microwave, proteinase K
H2O2 30 min 25 min 30 min 15 min
Primer antibody application, time and temperature 1 h, room temperature 1 h, room temperature 30 min, room temperature 1 h, 37°C
3,3’-diaminobenzidine tetrahydrochloride (DAB) 10 min 10 min 7 min 3 min
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