Production, purification and characterization of phytase from phytase producing fungus

Adnan Menderes Üniversitesi Fen-Edebiyat Fakültesi Biyoloji Bölümü Moleküler Biyoloji Anabilim Dalı stok kültürlerinde kayıtlı havadan ve topraktan izole edilmiş 165 adet fungus fitaz enzimi üretme kapasitesi açısından taranmıştır. Aspergillus niger UA-D suşunun en yüksek aktiviteye sahip olduğu saptanmıştır. Aspergillus niger UA-D suşundan ekstraselüler olarak PSM ortamında üretilen fitaz enzimi amonyum sülfat çöktürmesi, DEAE Sefaroz CL-6B ve Fenil Sepharoz CL-4B işlemleri uygulanarak % 2.6 verimle 73 kat saflaştırılmıştır. SDS-PAGE elektroforezinde tek bant elde edilmiş ve moleküler ağırlığı 65 kDa olarak hesaplanmıştır. Maksimum aktivitesini pH 3.0 ve 70 oC sergilediği bulunmuştur. Enzimin geniş bir sıcaklık ve pH aralığında stabil olduğu saptanmıştır. Enzimin fitik asit için Km değeri 180 µM ve Vmax değeri 54.35 U/mL olarak hesaplanmıştır. Fitaz enziminin geniş bir substrat spesifikliğine sahip olduğu belirlenmiştir. Fitaz aktivitesi, Ba2+ ve Li+ tarafından artarken, Fe2+, Al3+ ve Pb2+ tarafından inhibe olmuştur. Enzim aktivitesi NBS, PMSF, DTNB, 2,3-bütandion varlığında oldukça inhibe olurken, CMC, DTT ve ß-merkaptoetanol ile aktive olmuştur. Bu sonuçlar enzimin katalitik merkezinde triptofan, sistein, serin ve arjinin rezidülerinin önemli rol oynadığı sonucuna varılmıştır. Polihidrik alkollerden gliserol ve sorbitolün sıcaklık stabilitesini artırdığı ve organik çözücülerin ise enzim aktivitesini çok fazla etkilemediği belirlenmiştir.Fungal strains that were isolated from air and soil and registered in Adnan Menderes University Department of Biology culture stocks were screened for phytase activity of 165 fungal strains. Aspergillus niger UA-D. strain showed the highest phytase activity. Extracellular phytase produced by Aspergillus niger in PSM was purified 73-fold with a recovery of % 2.6 referred to the phytase activity in the crude extract using, ammonium sulphate precipitation, DEAE Sepharose CL-6B and Phenyl Sepharose CL-4B. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme gave a single stained band at a molecular mass of approximately 65 kDa. The temperature and pH for maximum activity of the enzyme were 70 oC and 3.0, respectively. Km and V max values for phytic acid of the enzyme were calculated to be, 180 µM and 54.35 U/mL, respectively. The phytase exhibited broad substrate specificity. The enzyme was stimulated by Ba2+ and Li+ and strongly inhibited by Fe2+, Al3+ and Pb2+ The enzyme activity was markedly inhibited in the presence of NBS, PMSF, DTNB, 2,3-butanedione suggesting that tryptophan, cysteine, arginine and serine residues play an important role in the catalytic process. Glyserol and sorbitol enhance thermostability of phytase. The enzyme was detemined to be highly stable against organic solvents

Ehrlichiosis ile doğal enfekte köpeklerde serum c-reaktive protein, prokalsitonin ve seruloplazmin konsantrasyonları

This study aimed to evaluate the concentrations of C-reactive protein (CRP), Procalcitonin (PCT), and ceruloplasmin (Cp), which are potential biochemical markers of the inflammatory process in dogs naturally infected with Ehrlichiosis. A total of 20 dogs, 8 clinically healthy (Healthy group) and 12 mono-infected with Ehrlichia spp. (Ehrlichia group) were included in the study. Dogs in the Ehrlichia group were selected from those showing clinical signs of active infection, and their diseases were diagnosed with commercial test kits. Serum CRP and PCT levels were analysed by commercially available test kits, and Cp concentration was determined by colourimetric methods. The CRP concentration in the Ehrlichia group was significantly higher compared to the healthy group. There was no significant difference between the groups in serum PCT and Cp concentrations. As a result, the increase in serum CRP concentration can be used for detecting inflammatory processes in dogs with Ehrlichiosis. In addition, this study showed that PCT and Cp concentrations are not clinically useful markers for determining inflammatory status in dogs with Ehrlichiosis.Bu çalışma, Ehrlichiosis ile doğal enfekte köpeklerde inflamatuar sürecin potansiyel biyokimyasal belirteçlerinden olan C-reaktif protein (CRP), Prokalsitonin (PCT) ve seruloplazmin (Cp) konsantrasyonlarını değerlendirmeyi amaçladı. Çalışmaya, klinik olarak sağlıklı 8 (Sağlıklı grup) ve Ehrlichia spp. ile mono enfekte 12 (Ehrlichia grubu) olmak üzere toplam 20 köpek dahil edildi. Ehrlichia grubundaki köpekler, klinik olarak aktif enfeksiyon belirtileri gösteren köpekler arasından seçildi ve hastalıkları, ticari test kitleri ile teşhis edildi. Serum CRP ve PCT seviyeleri köpek spesifik ticari ELISA test kitleri ile analiz edildi ve Cp konsantrasyonu kolorimetrik yöntemle belirlendi. Ehrlichia grubundaki CRP konsantrasyonu, sağlıklı grupla karşılaştırıldığında anlamlı olarak daha yüksekti. Serum PCT ve Cp konsantrasyonlarında gruplar arasında anlamlı fark yoktu. Sonuç olarak, serum CRP konsantrasyonundaki artış, Ehrlichiosis'li köpeklerde inflamatuar süreçlerin saptanmasında kullanılabilir. Ek olarak bu çalışma, PCT ve Cp konsantrasyonlarının Ehrlichiosis'li köpeklerde inflamatuar durumu belirlemek için klinik olarak yararlı belirteçler olmadığını göstermiştir

Histopathologic Evaluation of Nonalcoholic Fatty Liver Disease in Hypothyroidism-Induced Rats

It is speculated that thyroid hormones may be involved in nonalcoholic fatty liver disease (NAFLD) pathogenesis. A literature scan, however, demonstrated conflicting results from studies investigating the relationship between hypothyroidism and NAFLD. Therefore, our study aims to evaluate NAFLD, from the histopathologic perspective, in hypothyroidism-induced rats. Wistar rats were divided into 2 groups: the experimental group consumed water containing methimazole 0.025% (MMI, Sigma, USA) for 12 weeks and the control group consumed tap water. At the end of week 12, serum glucose, ALT, AST, triglyceride, HDL, LDL, TSH, fT4, fT3, visfatin, and insulin assays were performed. Sections were stained with hematoxylin-eosin and "Oil Red-O" for histopathologic examination of the livers. In our study, we detected mild hepatosteatosis in all hypothyroidism-induced rats. There was statistically significant difference with respect to obesity between the two groups ( < 0.001). The mean fasting blood glucose was 126.25 ± 23.4 mg/dL in hypothyroidism-induced group and 102.63 ± 15.51 mg/dL in the control group, with a statistically significant difference between the groups ( = 0.032). The two groups did not differ statistically significantly with respect to visfatin levels ( > 0.05). In conclusion, we found that hypothyroidism-induced rats had mild hepatosteatosis as opposed to the control group histopathologically. Our study indicates that hypothyroidism can cause NAFLD

Histopathologic Evaluation of Nonalcoholic Fatty Liver Disease in Hypothyroidism-Induced Rats

It is speculated that thyroid hormones may be involved in nonalcoholic fatty liver disease (NAFLD) pathogenesis. A literature scan, however, demonstrated conflicting results from studies investigating the relationship between hypothyroidism and NAFLD. Therefore, our study aims to evaluate NAFLD, from the histopathologic perspective, in hypothyroidism-induced rats. Wistar rats were divided into 2 groups: the experimental group consumed water containing methimazole 0.025% (MMI, Sigma, USA) for 12 weeks and the control group consumed tap water. At the end of week 12, serum glucose, ALT, AST, triglyceride, HDL, LDL, TSH, fT4, fT3, visfatin, and insulin assays were performed. Sections were stained with hematoxylin-eosin and “Oil Red-O” for histopathologic examination of the livers. In our study, we detected mild hepatosteatosis in all hypothyroidism-induced rats. There was statistically significant difference with respect to obesity between the two groups (p<0.001). The mean fasting blood glucose was 126.25 ± 23.4 mg/dL in hypothyroidism-induced group and 102.63 ± 15.51 mg/dL in the control group, with a statistically significant difference between the groups (p=0.032). The two groups did not differ statistically significantly with respect to visfatin levels (p>0.05). In conclusion, we found that hypothyroidism-induced rats had mild hepatosteatosis as opposed to the control group histopathologically. Our study indicates that hypothyroidism can cause NAFLD

CYTOTOXIC AND APOPTOTIC EFFECTS OF ZOLEDRONIC ACID IN D-17 CANINE OSTEOSARCOMA CELL LINE

ZOLEDRONİK ASİDİN D-17 KÖPEK OSTEOSARKOMA HÜCRE HATTINDA SİTOTOKSİK VE APOPTOTİK ETKİLERİ AŞICI EKREN G. S. Aydın Adnan Menderes Üniversitesi Sağlık Bilimleri Enstitüsü Biyokimya (Veteriner) Programı Doktora Tezi, Aydın, 2019. Çalışmamızda; in vitro olarak zoledronik asidin (ZA) D-17 (CCL-183) köpek osteosarkoma (OSA) hücre hattı üzerinde antiproliferatif etkisinin olup olmadığı ve olası hücre ölüm tipinin (apoptoz/nekroz) belirlenmesi amaçlandı. D-17 köpek osteosarkoma hücre hattına uygulanan zoledronik asidin (1 µM, 5 µM, 10 µM, 25 µM, 50 µM, 75 µM, 100 µM) hücre canlılığına etkisi ve IC50 değeri WST-1 canlılık testi ile 24, 48, 72 ve 96 saat inkübasyon sürelerince tespit edildi. WST-1 canlılık analizleri sonuçlarına göre 24 saatte anlamlı bir sitotoksik etki görülmezken (p≥0,05), ZA’nın IC50 değerleri 48. saat, 72. saat ve 96. saat için sırası ile 82,5 µM, 26,0 µM ve 17,6 µM olarak hesaplandı. ZA’nın hücrelerin koloni oluşturma yeteneklerine etkisi kristal violet, mineral içeriği üzerine etkisi alizarin kırmızısı boyaması ile gösterilirken, ZA’nın ekstraselüler ve intraselüler alkalen fosfataz aktivitesine olan etkisi ise spektrofotometrik yöntemle ölçüldü. ZA’nın hücrelerin koloni oluşturma, mineral içeriği ve alkalen fosfataz üzerine etkisi incelendiğinde hücrede doz ve zamana bağlı olarak azalmaya neden olduğu belirlendi (p<0,05). Hücrelerin göç etme potansiyellerine olan etkisi ucuz ve basit bir yöntem olan yara iyileştirme yöntemi ile belirlendi ve ZA’nın köpek OSA hücre hattı üzerine 5 µM, 7,5 µM, 10 µM, 15 µM konsantrasyonları uygulandığında hücrenin göç etme potansiyelini anlamlı ölçüde azalttığı görüldü. Zoledronik asidin hücredeki apoptotik etkileri ise elektroforetik ve ELİSA yöntemleri ile DNA kırıklarının analizi yapılarak gösterildi. Kaspaz 3, kaspaz 8 ve kaspaz 9 düzeyleri ELİSA yöntemi ile belirlenerek apoptotik yolaklardaki kaspaz-kaskat sistemi üzerine etkisi saptandı. Ayrıca apoptotik yolaklar üzerinde yer alan Bcl-2, Bax ve Bid genleri, hücrelerinin çoğaltılması ve sağkalımı ile ilişkili gen olarak kabul edilen survivin genlerinin ekspresyon düzeyleri RT-PCR yöntemleri kullanılarak belirlendi. Apoptotik etkilerinin saptanmasında WST-1 canlılık testi sonucu bulunan IC50 değerinin altındaki 5 µM, 7,5 µM, 10 µM, 15 µM konsantrasyonları kullanıldı. D-17 köpek OSA hücreleri üzerine seçilen dozlarda ZA uygulandığında doz ve zamana bağlı olarak apoptotik DNA kırıklarında, kaspaz 3 ve kaspaz 9 düzeyinde, Bax/Bcl-2 oranında anlamlı artış (p<0,05) gözlenirken, kaspaz 8 düzeyinde ve Bid ekspresyonunda anlamlı bir değişiklik bulunmadı (p≥0,05). Hücre sağkalımını belirleyen survivinin ekspresyon düzeylerinde ise doz ve zamana bağlı olarak anlamlı bir azalma (p<0,05) saptandı. Çalışmanın sonuçları ZA’nın D-17 köpek osteosarkoma hücrelerinde tek başına uygulanmasının in vitro olarak antikanserojenik etki gösterdiğini ortaya çıkarmıştır. Bu konudaki araştırmaların kapsamı genişletilmeli ve daha ileri moleküler çalışmalar gerçekleştirilmelidir. Ayrıca in vivo hayvan modelleri üzerinde denenerek doğrulukları kanıtlanmalıdır.CYTOTOXIC AND APOPTOTIC EFFECTS OF ZOLEDRONIC ACID IN D-17 CANINE OSTEOSARCOMA CELL LINE AŞICI EKREN G. S. Aydın Adnan Menderes University Health Sciences Institute Veterinary Biochemistry Program PhD Thesis, Aydın, 2019. The aim of our study was to determine whether zoledronic acid (ZA) has antiproliferative effect on D-17 (CCL-183) canine osteosarcoma (OSA) cell line in vitro and to determine possible cell death type (apoptosis/necrosis). The effect of zoledronic acid (1 μM, 5μM, 10 μM, 25 μM, 50 μM, 75 μM, 100 μM) on cell viability of the D-17 canine osteosarcoma cell line and the IC50 value were determined during 24, 48, 72 and 96 hours incubation time with the WST-1 viability test. According to the results of WST-1 vitality analysis, no significant cytotoxic effect was observed in 24 hours (p≥0,05), the IC50 values of ZA were calculated as 82,5 µM, 26,0 µM and 17,6 µM respectively for 48 th hour, 72 th and 96 th hours. The effect of zoledronic acid on the colony forming ability of the cells was shown by crystal violet, on mineral content with alizarin red staining while the effect of ZA on extracellular and intracellular alkaline phosphatase activity was measured by spectrophotometric method. When the effect of ZA on colony formation, mineral content and alkaline phosphatase was investigated, ıt was determined decreased on cell dependent on dose and time (p<0,05). The effect of cells on migration potentials was determined by a cheap and simple method is wound healing and ZA significantly decreased cell migration potential when the concentrations of 5 μM, 7,5 μM, 10 μM, 15 μM were applied to on the canine OSA cell line. Apoptotic effects of zoledronic acid in the cell were showed analysis by electrophoretic and ELISA methods. The effect on caspase-cascade system in the apoptotic pathways was detected by determining the caspase 3, caspase 8 and caspase 9 activities with ELISA method. Moreover, expression levels of survivin, regarded as genes related to proliferation and survival of Bcl-2, Bax and Bid genes and cells in the apoptotic pathways, were determined using RT-PCR methods. Below the IC50 value that found by the WST-1 viability test 5 µM, 7,5 µM, 10 µM, 15 µM concentrations were used to determine the apoptotic effects. When the selected doses of ZA were applied on D-17 canine osteosarcoma cells was significantly increased apoptotic DNA fragments, level of caspase 3 and caspase 9, Bax/Bcl-2 ratio (p<0,05) while there was no significant change in caspase 8 level and Bid expression (p≥0,05) depending on dose and time. There was a significant decrease (p<0,05) in the expression levels of the survivin which determined cell survival. The result of this study has been shown that the treatment of ZA alone in D-17 canine osteosarcoma cells shows an anticarcinogenic effect in vitro. The scope of research on this subject should be expanded and proven by testing on more advanced molecular studies and in vivo animal models.İÇİNDEKİLER KABUL VE ONAY SAYFASI i TEŞEKKÜR ii İÇİNDEKİLER iii SİMGELER VE KISALTMALAR DİZİNİ vii ŞEKİLLER DİZİNİ ix RESİMLER DİZİNİ xii TABLOLAR DİZİNİ xiii ÖZET xiv ABSTRACT xvi 1. GİRİŞ 1 2. GENEL BİLGİLER 4 2.1. Kanser 4 2.2. Hücre Ölümü 4 2.3. Programlanmış Hücre Ölümü ve Tipleri 5 2.4. Apoptoz 7 2.5. Apoptozun Keşfi ve Tarihçesi 9 2.6. Apoptoz ve Nekroz 11 2.7. Apoptotik Süreç 15 2.8. Apoptozun Biyokimyası 16 2.9. Apoptozun Uyarılması 18 2.10. Apoptozun Etki Mekanizması 19 2.10.1. Ekstrinsik Yolak 19 2.10.1.1. Fas-Fas ligand aracılı apoptoz (CD95 Yolu) 21 2.10.1.2. Tümor nekroz faktör aracılı apoptoz 22 2.10.2. İntrinsik Yolak 23 2.10.3. Sitotoksik T Lenfosit Aracılı Apoptoz (Perforin/Granzim Yolu) 24 2.10.4. Endoplazmik Retikulum Aracılı Apoptoz 26 2.10.5. Kaspaz Bağımsız Apoptoz 28 2.11. Kaspazlar 29 2.11.1. Kaspazların Yapısı 32 2.11.2. Kaspaz 3 34 2.11.3. Kaspaz 8 35 2.11.4. Kaspaz 9 36 2.12. Bcl-2 Gen Ailesi 36 2.13. Survivin (BIRC5) 39 2.14. Köpek Osteosarkoma 42 2.15. Bisfosfonatlar 46 2.15.1. Bisfosfonatların Tarihçesi 46 2.15.2. Bisfosfonatların Yapısı 47 2.15.3. Bisfosfonatların Metabolik Etkileri 49 2.16. Zoledronik Asit 51 3. GEREÇ VE YÖNTEM 57 3.1. Gereç 57 3.1.1. Hücre Hattı 57 3.1.2. Kullanılan Kimyasal Maddeler 57 3.1.3. Kullanılan Sarf Malzemeler 57 3.1.4. Kullanılan Cihazlar 58 3.1.5. Kullanılan Kitler 58 3.1.6. Primerler 58 3.1.7. Kullanılan Besiyeri ve Çözeltilerin Hazırlanışı 61 3.2. Yöntem 62 3.2.1. Sterilizasyon 62 3.2.2. D-17 (CCL-183) Köpek Osteosarkoma Hücre Hattının Kültüre Edilmesi 63 3.2.3. Hücrelerin Dondurulması ve Açma İşlemi 64 3.2.4. Hücrelerin Kaplama Yoğunluğunun Belirlenmesi 65 3.2.5. Zoledronik Asidin Doz ve Zamana Bağlı Olarak Hücre Canlılığı Üzerine Etkisi 65 3.2.6. WST-1 (Water Soluble Tetrazolium tuzu) Canlılık Testi 66 3.2.7. IC50 Değerinin Hesaplanması 67 3.2.8. Koloni Oluşturabilme Yeteneklerinin İncelenmesi 67 3.2.9. Zoledronik Asidin Mineralizasyona Etkisi 68 3.2.10. Zoledronik Asidin Alkalen Fosfataz (ALP) Aktivitesine Olan Etkisi 68 3.2.11. Zoledronik Asidin Hücrelerin Migrasyonuna Olan Etkileri 69 3.2.12. Zoledronik Asidin D-17 (CCL-183) Köpek Osteosarkoma Hücreleri Üzerinde Apoptotik Etkileri 70 3.2.12.1. Apoptotik DNA fragmentlerinin ELİSA yöntemi ile kantitatif analizi 70 3.2.12.2. Apoptotik DNA fragmentasyonun ekstraksiyonu ve agaroz jel elektroforezinde görüntülenmesi 71 3.2.13. Kaspaz 3, Kaspaz 8 ve Kaspaz 9’nın ELİSA ile Belirlenerek Apoptozun Değerlendirilmesi 71 3.2.14. RT-PCR ile Gen İfadelerinin Belirlenmesi 73 3.2.14.1. Total RNA izolasyonu 73 3.2.14.2. cDNA sentezi 74 3.2.14.3. Primerlerin hazırlanışı 74 3.2.14.4. mRNA ekspresyon düzeylerinin qRT-PCR yöntemi ile analizi 75 3.2.14.5. Ekspresyonun değerlendirilmesi 76 3.2.14.6. PCR ürünlerinin agaroz jel elektroforezinde görüntülenmesi 77 3.2.15. İstatistiksel Analiz 77 4. BULGULAR 78 4.1. Hücrelerin Kaplama Yoğunluğunun Belirlenmesi 78 4.2. Zoledronik Asidin Hücre Canlılığı Üzerine Etkisi 79 4.3. Zoledronik Asidin D-17 Köpek Osteosarkoma Hücre Hattı Üzerine Koloni Oluşturabilme Yeteneklerine Etkisi 83 4.4. Zoledronik Asidin Mineralizasyona Etkisi 85 4.5. Zoledronik Asidin Alkalen Fosfataz (ALP) Aktivitesine Olan Etkisi 85 4.6. Zoledronik Asidin Hücrelerin Migrasyonuna Olan Etkisi 88 4.7. Zoledronik Asidin D-17 (CCL-183) Köpek Osteosarkoma Hücreleri Üzerinde Apoptotik Etkileri 91 4.7.1. Apoptotik DNA Fragmentlerinin ELİSA Yöntemi ile Kantitatif Analizi 91 4.7.2. Apoptotik DNA Fragmentasyonun Ekstraksiyonu ve Agaroz Jel Elektroforezinde Görüntülenmesi 93 4.7.3. D-17 (CCL-183) Köpek Osteosarkoma Hücre Hattında Zoledronik Asidin Kaspaz 3, 8, 9 Aktivitesine Olan Etkileri 94 4.7.4. Genlerin Kantitatif Ekspresyon Düzeylerinin Belirlenmesi 97 4.7.4.1. Melting point analizi 100 4.7.4.2. PCR ürününün agaroz jel elektroforezinde görüntülenmesi 102 5. TARTIŞMA 103 6. SONUÇ VE ÖNERİLER 113 KAYNAKLAR 115 ÖZGEÇMİŞ 15

Cytotoxic and apoptotic effect of nanoclinoptilolite on canine osteosarcoma cell lines

Clinoptilolite has antiviral, antibacterial, anti-inflammatory, antidiabetic, and anticancer properties due to its biological activities. In various cancer cell culture studies, it has been reported effective against tumour cells and gave positive results in treatment of various tumours in dogs. No study was found on the effects of the nanoparticulate form, nanoclinoptilolite, on cancer cells. The aim of this study was to determine its cytotoxic and apoptotic effects in canine osteosarcoma (OSA) cell culture

Effects of the season on physiological and endocrine traits and on HSP70 in Saanen goats under Mediterranean climate conditions

WOS: 000419447900006This study was conducted to determine the impact of heat stress on some physiological and endocrine traits in Saanen goats raised under Mediterranean climate conditions. The effects of thermal stress on heart rate (HR), respiration rate (RR) and rectal temperature (RT) on plasma total trii-odothyronine (T3), thyroxine (T4), Cortisol (C), and HSP70 concentrations were evaluated on twenty two Saanen goats of different ages in the second week of April 2013, July 2013, October 2013 and January 2014. Climatic data such as temperature (degrees C) and relative humidity (%) were recorded from Spring 2013 to Winter 2014. Live body and BCS values were also recorded during this period. The physiological parameters above were measured twice on each experiment day (morning and afternoon) in all seasons. Blood samples were collected in each afternoon of the experiment day to analyze T3, T4 C, and HSP70. All data were analyzed. According to the values of rectal temperature (RT), it was estimated that the goats were under extreme heat stress only in the summer season. The heart rate (HR) values in the winter season for morning and noon periods were found statistically significant (p < 0.05). The average respiratory rate (RR) in the spring season was found significantly lower. On the other hand, the RR for the noon period in the summer was higher than in the fall and winter seasons (p < 0.05). There was a significant difference (p < 0.05) between summer and fall seasons for C values. The highest value (96.62 ng/ml) was obtained in spring, whereas the lowest (60.58 ng/ml) in the fall. T3 levels in the fall and winter were found to be statistically significant (p < 0.05). They were the highest in the winter and spring, and the lowest in fall. T4 and T4/T3 levels in the winter were found to be statistically higher than in other seasons (p < 0.05). The lowest value for T4 was found in the fall and for T4/T3 in summer. Mean HSP70 value in spring was found to be statistically low (p < 0.05). The changes in THI values in different seasons, particularly between mornings and afternoons, indicated that thermal stress was evident, and that the animals became resistant to it eventually. The fluctuations of the C, T3, T4 and HSP70 values were indicators of the animals' reaction to thermal stress. The THI values in spring, which were between 16 and 18 THI, may be considered within the ideal comfort zone for goats. It was observed that Saanen goats were able to adapt to seasonal weather changes in the environmental conditions of the region