dc.contributor.advisor |
Eren, Ülker |
|
dc.contributor.author |
Bozkurt Di Stefano, Müge |
|
dc.date.accessioned |
2022-07-19T06:30:32Z |
|
dc.date.available |
2022-07-19T06:30:32Z |
|
dc.date.issued |
2022 |
|
dc.date.submitted |
2022 |
|
dc.identifier.citation |
Bozkurt Di Stefano, M. (2022) Farklı yaşam ortamında bulunan balıkların kan hücreleri üzerinde morfolojik ve sitokimyasal çalışmalar (basılmamış Doktora Tezi) Aydın Adnan Menderes Üniversitesi Sağlık Bilimleri Enstitüsü, Aydın |
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dc.identifier.uri |
http://hdl.handle.net/11607/4673 |
|
dc.description.abstract |
Amaç: Sunulan çalışmada, tuzlu su balıklarından olan Deniz levreği ve Çipura ile tatlı su
balığı olan Gökkuşağı alabalığında, perifer kan hücrelerinin morfolojilerinin ve sitokimyasal
özelliklerinin incelenmesi amaçlanmıştır.
Gereç ve Yöntem: Çalışmada; Deniz levreği, Çipura ve Gökkuşağı alabalığı olmak üzere, her
türden sekiz adet balıktan, heparin ve EDTA’lı tüplere alınan kan örnekleri, materyal olarak
kullanılmıştır. Hazırlanan frotilere; CR, PAS, SB, TB, MGG, MGP, ANAE ve sABC boyama
metotları uygulanıp, ışık mikroskobik düzeyde inceleme gerçekleştirilmiştir.
Bulgular: Uygulanan sitokimyasal metotlarla, her üç balık türünde de memeli ve kanatlılara
benzer şekilde kan hücreleri tespit edilmiştir. Eritrosit, lenfosit ve trombosit ayırımında sorun
yaşanmamıştır. Granülositler açısından ise balık türlerine göre hücrelerin, sitokimyasal
metotlardan önce kullanılan antikoagulan tipinden, bazen de tespit işleminden etkilenebildiği
belirlenmiştir. Granülositlerde CR, SB, PAS, TB, MGG, MGP pozitivitelerinin balık türlerine
göre özellikleri ve antikoagulandan etkilenme durumları kaydedilmiştir. Enzimhistokimyasal
olarak, balık türlerine göre değişmekle beraber lenfosit, monosit ve granülositlerde
nonspesifik esteraz (ANAE) pozitivitesi belirlenmiştir. Ayrıca, immunhistokimyasal olarak da
Deniz levreğinde DLT15 monoklonal antikoru kullanarak T lenfositler tespit edilmiştir.
Sonuç: Balık kanında sitokimyasal çalışmalar yapılırken antikoagulan kullanımı ve tespit
işlemi uygulanmasında farklı alternatiflerin denenmesi gerektiği sonucuna varılmıştır.
Hücrelerinin morfolojik ve sitokimyasal özelliklerinin, yaşam ortamı farklılığından ziyade,
taksonomik olarak familya farklılığından kaynaklandığı düşünülmüştür. Sunulan çalışmada
elde edilen bilgiler temel alınarak, granülositlerin granül içeriklerinin, enzimhistokimyasal
yöntemlerle, ışık ve elektron mikroskobik düzeyde incelenmesi, nonspesifik ve spesifik
immunitenin evrimsel gelişiminin öğrenilmesine katkıda bulunacaktır. |
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dc.description.abstract |
Objective: In the present study, it was aimed to investigate the morphology and cytochemical
properties of peripheral blood cells in sea bass and sea bream, which are saltwater fish, and
rainbow trout, which is freshwater fish.
Materials and Methods: In the study; blood samples taken from eight fish of each species,
including sea bass, sea bream and rainbow trout, into tubes with heparin and EDTA were used
as material. To the prepared smears; CR, PAS, SB, TB, MGG, MGP, ANAE and sABC
staining methods were applied and light microscopic examination was performed.
Results: Based on cytochemical results, blood cells were detected in all three fish species,
similar to mammals and birds. There was no problem in the differentiation of erythrocyte,
lymphocyte and thrombocyte. In terms of granulocytes, it was determined that the cells could
be affected by the type of anticoagulant used before the cytochemical methods, and
sometimes by the fixation process, depending on the fish species. The characteristics of CR,
SB, PAS, TB, MGG, MGP positivity in granulocytes according to fish species and their
response to anticoagulant were recorded. Enzymhistochemically, nonspecific esterase
(ANAE) positivity was determined in lymphocytes, monocytes and granulocytes, although it
varies according to fish species. In addition, T lymphocytes were detected
immunohistochemically in sea bass using DLT15 monoclonal antibody.
Conclusion: It has been concluded that different alternatives should be tried in the use of
anticoagulants and the application of fixation while performing cytochemical studies in fish
blood. It was thought that the morphological and cytochemical characteristics of the cells
were caused by the taxonomic family difference rather than habitat differences. In the light of
the information obtained in the presented study, the examination of the granule contents of
granulocytes by enzymehistochemical methods, at the light and electron microscopic level
will contribute to the learning of the evolutionary development of nonspecific and specific
immunity. |
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dc.description.tableofcontents |
İÇİNDEKİLER
KABUL VE ONAY…………………………………………………………............ i
TEŞEKKÜR………………………………………………………………………… ii
İÇİNDEKİLER……………………………………………………………………… iii
SİMGELER VE KISALTMALAR DİZİNİ………………………………………… vii
ŞEKİLLER DİZİNİ…………………………………………………………………. ix
RESİMLER DİZİNİ………………………………………………………………… x
TABLOLAR DİZİNİ……………………………………………………………….. xvii
ÖZET………………………………………………………………………………... xviii
ABSTRACT…………………………………………………………………………. xix
1. GİRİŞ……………………………………………………………………………… 1
1.1. Balık Filogenisi…………………………………………………………………. 2
1.2. Balıklarda Yaşam Ortamı ve Beslenme Farklılığı……………………………… 3
1.3. Balık Biyolojileri………………………………………………………………… 5
1.3.1. Deniz Levreği (Dicentrarchus labrax L., 1758) Biyolojisi……………………. 5
1.3.2. Çipura (Sparus aurata L., 1758) Biyolojisi……………………………………. 7
1.3.3. Gökkuşağı Alabalığı (Oncorhynchus mykiss Walbaum, 1792) Biyolojisi…….. 9
1.4. Balıklarda Perifer Kan Hücrelerinin Memelilere Göre Farklı Özellikleri……….. 11
2. GENEL BİLGİLER………………………………………………………………… 12
2.1. Balıklarda Kan Hücreleri…………………………………………………………. 12
2.1.1. Eritrositler………………………………………………………………………. 13
2.1.2. Lökositler……………………………………………………………………….. 14
2.1.2.1. Lenfosit Lökositler…………………………………………………………….. 14
iv
2.1.2.2. Monosit Lökositler ……………………………………………………………....... 15
2.1.2.3. Nötrofil Lökositler …………………………………………………………............ 16
2.1.2.4. Eozinofil Lökositler ……………………………………………………………….. 16
2.1.2.5. Bazofil Lökositler ……………………………………………………………….... 17
2.1.3. Trombositler ve Hemostaz…………………………………………………………. 18
2.2. Hematopoezis………………………………………………………………………… 18
2.3. Perifer Kan Hücreleri ile İlgili Sitokimyasal ve Enzimhistokimyasal Boyama
Metodları…………………………………………………………………………….. 20
2.3.1. May-Grünwald-Giemsa (MGG) Boyama Metodu…………………………………. 20
2.3.2. Congo Red (CR) Boyama Metodu………………………………………………….. 21
2.3.3. Periodic Acid Shiff (PAS) Boyama Metodu………………………………………... 21
2.3.4. Sudan Black (SB) Boyama Metodu………………………………………………… 22
2.3.5. Toluidine Blue (TB) Boyama Metodu……………………………………………… 22
2.3.6. Methyl Green Pyronin (MGP) Boyama Metodu…………………………………… 23
2.3.7. Alpha Naphthyl Acetate Esterase (ANAE) Boyama Metodu……………………… 23
3. GEREÇ ve YÖNTEM………………………………………………………………...... 25
3.1. Gereç…………………………………………………………………………………. 25
3.2. Yöntem……………………………………………………………………………….. 29
3.2.1. MGG Boyama Metodunun Uygulanması………………………………………….. 30
3.2.2. CR Boyama Metodunun Uygulanması……………………………………………... 30
3.2.3. PAS Boyama Metodunun Uygulanması…………………………………………..... 30
3.2.4. SB Boyama Metodunun Uygulanması……………………………………………... 31
3.2.5. TB Boyama Metodunun Uygulanması…………………………………………....... 31
3.2.6. MGP Boyama Metodunun Uygulanması………………………................................ 32
v
3.2.7. ANAE (Ph: 5.8) Boyama Metodunun Uygulanması……………………………….. 32
3.2.8. Streptavidin-Biotin-Peroksidaz Kompleks (sABC) Boyama Metodunun
Uygulanması……………………………………………………………………….. 33
4. BULGULAR…………………………………………………………………………… 35
4.1. Deniz Levreği Kanı…………………………………………………………………... 37
4.1.1. CR Boyama Metodu……………………………………………………................... 37
4.1.2. PAS Boyama Metodu………………………………………………………………. 38
4.1.3. SB Boyama Metodu………………………………………………………………... 44
1. TB Boyama Metodu……………………………………………………………….......... 45
4.1.5. MGG Boyama Metodu…………………………………………............................... 47
4.1.6. ANAE PH 5.8 Boyama Metodu……………………………………………………. 51
4.1.7. sABC Boyama Metodu…………………………………………………………….. 51
4.1.8. MGP Boyama Metodu……………………………………………………………… 52
4.2. Çipura Kanı…………………………………………………………………………... 53
4.2.1. CR Boyama Metodu……………………………………………………................... 53
4.2.2. PAS Boyama Metodu………………………………………………………………. 56
4.2.3. SB Boyama Metodu………………………………………………………………... 59
4.2.4. TB Boyama Metodu………………………………………………………………... 62
4.2.5. MGG Boyama Metodu…………………………………………............................... 62
4.2.6. ANAE PH 5.8 Boyama Metodu……………………………………………………. 69
4.2.7. MGP Boyama Metodu……………………………………………………………… 70
4.3. Gökkuşağı Alabalığı Kanı…………………………………………………………… 72
4.3.1. CR Boyama Metodu………………………………………………………………... 72
4.3.2. PAS Boyama Metodu………………………………………………………………. 73
4.3.3. SB Boyama Metodu……………………………………………………………….... 76
vi
4.3.4. TB Boyama Metodu…………………………………………………...................... 77
4.3.5. MGG Boyama Metodu…………………………………………............................... 78
4.3.6. ANAE PH 5.8 Boyama Metodu……………………………………………………. 83
4.3.7. MGP Boyama Metodu……………………………………………………………... 85
5. TARTIŞMA…………………………………………………………………………..... 87
5.1. CR Boyama Metodu………………………………………………………………..... 88
5.2. PAS Boyama Metodu………………………………………………………………... 89
5.3. SB Boyama Metodu………………………………………………………………….. 91
5.4. TB Boyama Metodu………………………………………………………………….. 92
5.5. MGG Boyama Metodu……………………………………………………………..... 93
5.6. ANAE Boyama Metodu……………………………………………………………... 95
5.7. MGP Boyama Metodu……………………………………………………………..… 96
6. SONUÇ ve ÖNERİLER………………………………………………………………... 97
KAYNAKLAR…………………………………………………………………………… 99
EKLER……………………………………………………………………………….…… 119
EK 1 (ADÜ-HADYEK)………………………………………………………………....... 119
BİLİMSEL ETİK BEYANI……………………………………………………………… 120
ÖZ GEÇMİŞ……………………………………………………………………………… 121 |
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dc.language.iso |
tur |
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dc.publisher |
Aydın Adnan Menderes Üniversitesi Sağlık Bilimleri Enstitüsü |
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dc.rights |
info:eu-repo/semantics/openAccess |
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dc.subject |
Antikoagulan, Çipura, Deniz levreği, Gökkuşağı alabalığı, Perifer kan |
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dc.subject |
Anticoagulant, Sea bream, Sea bass, Rainbow trout, Peripheral blood |
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dc.title |
Farklı yaşam ortamında bulunan balıkların kan hücreleri üzerinde morfolojik ve sitokimyasal çalışmalar |
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dc.title.alternative |
Morphological and cytochemical studies on blood cells of fish in different habitats |
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dc.type |
doctoralThesis |
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dc.contributor.department |
Aydın Adnan Menderes Üniversitesi Sağlık Bilimleri Enstitüsü |
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