Isolation and characterization of bacillus thuringiensis strains from different grain habitats

Thesis (Master)--Izmir Institute of Technology, Biotechnology, Izmir, 2004Includes bibliographical references (leaves: 46-58)Text in English; Abstract: Turkish and Englishvii, 58 leavesBacillus thuringiensis is a Gram positive, facultative anaerob bacteria that produces proteins toxic against different insect species. This feature makes it the most widely used biological control agent in agriculture. Since B. thuringiensis strains have great genetic diversity, the toxic behaviours of these strains differ from region to region. Native B. thuringiensis strains are isolated from different habitats and characterized to determine their toxic potential all over the world. The aim of this study was to isolate B. thuringiensis strains from different grain habitats in Central Anatolia and Aegean Regions, and to investigate their phenotypic and genotypic characterizations. Total 96 samples containing soil, grain, stored product dust, straw and various residues were collected from wheat farms, grain silos, haylofts and caves in Ereli/Konya, Takale/Karaman, Nikfer/Denizli, and Bozbük/Söke under aseptic conditions. Seven hundred bacteria were isolated from these samples by sodium acetate selection and heat treatment. For phenotypic characterization, 500 of these isolates were grown for 48 h and crystal protein production was observed by phase contrast microscobe during spore formation. One hundred and sixty three of the bacterial colonies were identified as B. thuringiensis. The isolates were divided into 5 different groups based on the shape of the crystals that they produced. Spherical type crystal morphology was mostly observed type among the others. For genotypic characterization, the cry gene content of the isolates were screened by polymerase chain reaction (PCR) analysis. In addition, chromosomal DNA analysis of 34 isolates by Pulsed Field Gel Electrophoresis (PFGE) as well as plasmid DNA profiling for all isolates were also carried out. One hundred and three isolates were positive for 5 different cry genes (cry1, cry2, cry3, cry4, cry9) examined by PCR. Among all cry genes examined, cry1 and cry9 genes were mostly found in the isolates. Morover, plasmid profiling of the isolates indicated that a 15 kb DNA band was present in all the isolates; however, some of them had more than one DNA band at different sizes. Finally, chromosomal DNA profiling by PFGE showed different DNA patterns for isolates containing the same cry gene which suggest a high level of diversity among the B. thuringiensis strains isolated. Further studies related with extensive genetic characterization and toxic activity of each B. thuringiensis strain will give more comprehensive results on biodiversity of B. thuringiensis strains in Anatolia

Isolation and characterization of Bacillus thuringiensis strains from different grain habitats in Turkey

Bacillus thuringiensis (Bt) is a gram-positive, spore-forming bacterium and it produces insecticidal crystal (cry) proteins during sporulation. Because the genetic diversity and toxic potential of Bt strains differ from region to region, strains have been collected and characterized all over the world. The aim of this study is to isolate Bt strains in grain-related habitats in Turkey and to characterize them on the basis of crystal morphology, cry gene content, and chromosomal and plasmid DNA profiles. Four approaches were taken analysis with phase contrast (PC) microscopy, polymerase chain reaction (PCR), pulsed field gel electrophoresis (PFGE) and plasmid isolation. Ninety-six samples were collected from Central Anatolia and the Aegean region. Bt was isolated from 61 of 96 samples (63.5) and 500 Bt-like colonies were obtained. One hundred and sixty three of the colonies were identified as Bt based on cry protein formation using PC microscopy. Among the examined colonies, the overall proportion identified (as Bt index) was 0.33. We found that 103 isolates were positive for the five different cry genes (cry1, cry2, cry3, cry4 and cry9) examined with PCR. In addition, plasmid profiling of 37 cry gene-positive isolates indicated that the 15 kb plasmid band was present in all isolates; however, 11 of 37 isolates had more than one plasmid band at different sizes. Finally, chromosomal DNA profiling by PFGE gave rise to different DNA patterns for isolates containing the same cry gene which suggests a high level of diversity among the Bt strains isolated.İYTE: 2002 IYTE43; DPT: 2002K-120739

Isolation and characterization of bacillus thuringiensis strains from different grain habitats

Thesis (Master)--Izmir Institute of Technology, Biotechnology, Izmir, 2004Includes bibliographical references (leaves: 46-58)Text in English; Abstract: Turkish and Englishvii, 58 leavesBacillus thuringiensis is a Gram positive, facultative anaerob bacteria that produces proteins toxic against different insect species. This feature makes it the most widely used biological control agent in agriculture. Since B. thuringiensis strains have great genetic diversity, the toxic behaviours of these strains differ from region to region. Native B. thuringiensis strains are isolated from different habitats and characterized to determine their toxic potential all over the world. The aim of this study was to isolate B. thuringiensis strains from different grain habitats in Central Anatolia and Aegean Regions, and to investigate their phenotypic and genotypic characterizations. Total 96 samples containing soil, grain, stored product dust, straw and various residues were collected from wheat farms, grain silos, haylofts and caves in Ereli/Konya, Takale/Karaman, Nikfer/Denizli, and Bozbük/Söke under aseptic conditions. Seven hundred bacteria were isolated from these samples by sodium acetate selection and heat treatment. For phenotypic characterization, 500 of these isolates were grown for 48 h and crystal protein production was observed by phase contrast microscobe during spore formation. One hundred and sixty three of the bacterial colonies were identified as B. thuringiensis. The isolates were divided into 5 different groups based on the shape of the crystals that they produced. Spherical type crystal morphology was mostly observed type among the others. For genotypic characterization, the cry gene content of the isolates were screened by polymerase chain reaction (PCR) analysis. In addition, chromosomal DNA analysis of 34 isolates by Pulsed Field Gel Electrophoresis (PFGE) as well as plasmid DNA profiling for all isolates were also carried out. One hundred and three isolates were positive for 5 different cry genes (cry1, cry2, cry3, cry4, cry9) examined by PCR. Among all cry genes examined, cry1 and cry9 genes were mostly found in the isolates. Morover, plasmid profiling of the isolates indicated that a 15 kb DNA band was present in all the isolates; however, some of them had more than one DNA band at different sizes. Finally, chromosomal DNA profiling by PFGE showed different DNA patterns for isolates containing the same cry gene which suggest a high level of diversity among the B. thuringiensis strains isolated. Further studies related with extensive genetic characterization and toxic activity of each B. thuringiensis strain will give more comprehensive results on biodiversity of B. thuringiensis strains in Anatolia

Türk Öğretmenlerinin Duygusal Zekâ ile ilgili görüşlerinin SEM ile analizi

The main purpose of this research is to identify the views of Turkish school teachers and principals with respect to the concept of emotional intelligence. A total of 169 questionnaires are evaluated using a modified version of the 'competence scale' of Titrek (2004). Explanatory factor analysis and confirmatory factor analysis are used to define the dimensions of emotional intelligence. The findings indicate no significant difference in views about emotional intelligence when comparing the gender, status, and living places of the teacher respondents, but there are significant differences when comparing their age and seniority.Bu çalışmanın amacı Türk öğretmen ve yöneticilerin duygusal zekâ ile ilgili görüşlerinin SEM analizi ile incelemektir. Araştırmaya Antalya, Burdur ve Isparta illerindeki ilköğretim okullarında görev 169 yapan öğretmen katılmıştır.   Araştırmada "Duygusal Zekâ'ya İlişkin Yeterlikler Ölçeği" (Titrek, 2004)  kullanılmıştır. Ölçeğin yazarından izin alınarak ölçek kullanılmıştır. Ölçek 72 maddeden oluşmaktadır. Ölçek Katılmıyorumdan (1)- Katılıyoruma (5) doğru giden beşli Likert tipi ölçek olarak düzenlenmiştir.Verilerin analizinde SPSS 11,5'den yararlanılmıştır. Cinsiyet değişkenleri için T- testi yapılmış; öğretmen ve yöneticilerin yaşları, kıdemleri, yaşadıkları şehirler, görevlerini içeren değişkenlere F testi ve Scheffée testi uygulanmıştır. Araştırmada duygusal zekayı etkileyen faktörlerin belirlenmesinde doğrulayıcı faktör analizi kullanılmıştır. Doğrulayıcı faktör analizi LISREL 8.54 ile gerçekleştirilmiştir.  Ölçek alan yazın taramasına dayalı olarak araştırma konusuna göre yeniden uyarlanmıştır. Ölçekte kaç faktör olması gerektiği ve maddelerden hangilerinin hangi faktörlerle üst düzeyde ilişkili olduklarını belirleyebilmek için açıklayıcı faktör analizi tekniği kullanılmıştır. Açıklayıcı faktör analizi sonucunda 24 maddenin 5 faktör (duyguları güdüleme, sosyal beceriler, duyguları yönetme, özbilinç ve empati)  altında toplanmasına karar verilmiştir. Yapılan analize göre sosyal beceriler faktörü en düşük ortalamaya, duyguları yönetme faktörü ise en yüksek ortalamaya sahip faktör olarak tespit edilmiştir. Sonuç olarak, duygusal zekayı etkileyen faktörlerden sosyal beceriler faktörü en düşük ortalamaya sahiptir ve duygusal zekayı etkilemede son sıralardadır. Doğrulayıcı faktör analizi sonuçlarına göre duygusal zekayı en fazla etkileyen  "empati", en az etkileyen boyut ise  "duyguları yönetme"'dir. Yapılan analizler sonucunda cinsiyetin duygusal zeka faktörlerini etkilemediği görülmektedir.  Öğretmenlerin yaşadıkları şehirlerin duygusal zeka üzerinde etkisi olmadığı bulunmuştur. Yaş ile duygusal zeka faktörleri arasında yapılan Anova testi sonucunda yaş ile duyguları güdüleme; duyguları yönetme ile özbilinç faktörleri arasında anlamlı farklılık görülmüştür.  Öğretmenlerin kıdemi arttıkça duyguları güdüleme ve duyguları yönetmeden etkilendikleri görülmektedir. Öğretmenlerin görevlerinin duygusal zeka üzerinde etkisi olmadığı tespit edilmiştir

Identification and bioactivity of native strains of Bacillus thuringiensis from grain-related habitats in Turkey

A native collection of Bacillus thuringiensis (Bt) strains originated from grain-related habitats in Turkey was characterized according to serotype, cry1 gene content, and bioactivity against Ephestia kuehniella (Lepidoptera: Phycitidae) and Spodoptera littoralis (Lepidoptera: Noctuidae). Twenty-three different serotypes as well as 24 unknown serotypes were obtained from 56 positively agglutinated strains with previously characterized antisera. Most common serovars were sotto, kim, and tochigiensis with the percentages of 14, 14, and 13, respectively. Among the cry1 gene-positive 36 strains, cry1E (100%), cry1Aa (94%), cry1Ac (92%), and cry1D (83%) genes were the most abundant. Bioactivity tests with 56 Bt strains carrying cry1, cry2, and/or cry9 genes indicated that all of them resulted in growth retardation or inhibition of larvae of both E. kuehniella and S. littoralis; however, only one strain, 85PPb (serovar morrisoni), caused high mortality in both insects (84% and 100%, respectively). Different crystal morphology was observed for the strain 85PPb and the standard strain B. thuringiensis subsp. morrisoni. Finally, no correlation was found among serotype, cry gene content and biotoxicity of Bt strains in the collection.State Planning Organization, DPT (2002K-120/390

Chronic venous insufficiency patterns in lower extremity veins detected by Doppler Ultrasound

Objective: The aim of this study was to reveal patterns ofchronic venous insufficiency of lower extremity detectedby color Doppler ultrasound and clarify its clinical implicationsin the patients with varicose veins symptoms.Methods: Between 2006 to 2011, a total of 2006 patientspresented with symptoms of venous insufficiency wereincluded in study. A total of 3938 lower extremity venoussystems were examined using color Doppler US. Weclassified four patterns considering combinations of superficialand deep venous insufficiency.Results: Of 2006 patients, 966 had either single venousinsufficiency (790, 82%) or combined insufficiency (176,18%). Superficial venous insufficiency was observedat the saphenoefemoral junction (SFJ) in 25.5%, at theGreat Saphenous Vein (GSV) in 57.6%, at the GiacominiVein in 2.4%, at the saphenopopliteal junction in 1.8% andfinally at the Small Saphenous Vein (SSV) in 9.1% of thepatients. We found multilevel venous insufficiency showingconnections at the rate of 51%. We described fourpatterns as Pattern 1: SFJ insufficiency combined withGSV (97.9%), GSV branching (7.1%), and perforatingvein (20.8%) insufficiency, Pattern 2: Deep venous insufficiencycombined with SFJ (63.6%), GSV (76.4%), andSSV (16.4%) insufficiency, Pattern 3: SPJ insufficiencycombined with SSV insufficiency (95.5%) and Pattern 4:Giacomini Vein insufficiency combined with GSV (67.9%)and SSV (75%) insufficiency.Conclusion: Chronic venous insufficiency may show fourdifferent patterns. Our results revealed that SFJ and GSVinsufficiency combination and deep venous insufficiencyand GSV insufficiency combinations are the most commoninsufficiency patterns seen in lower extremity.Key words: Doppler ultrasound, varicose veins, venous insufficienc

Isolation and characterization of Bacillus thuringiensis strains from olive-related habitats in Turkey

Aims: To isolate Bacillus thuringiensis strains from different olive-related habitats (olive groves and olive oil factories) in Turkey and to characterize these strains by molecular methods. Methods and Results: A total of 150 samples, consisting of olive grove soil, green olive leaves, olive leaf residues, animal faeces, olive pomace and dust, were examined for the presence of B. thuringiensis. One hundred B. thuringiensis strains were isolated from 54 environmental samples (36%) and characterized in terms of crystal morphology, cry and cyt gene content by polymerase chain reaction, plasmid profiles and 16S-internal transcribed spacer ribosomal DNA restriction fragment length polymorphism (16S-ITS rDNA RFLP). The highest percentage of samples containing B. thuringiensis was found in 38 out of 54 total soil samples (70%). Of the 100 B. thuringiensis isolates, the most frequent crystal shapes were irregularly shaped (24%), spherical-irregular pointed (19%), cuboidal (17%) and spherical (16%). The cry1 plus cry4 genotype was the most abundant genotype in our collection (21%). RFLP analysis of the amplified 16S-ITS rDNA revealed 11 distinct patterns for the isolates and 10 reference strains. Conclusions: Bacillus thuringiensis isolates showed a great genetic diversity and crystal shape heterogeneity. Significance and Impact of the Study: This is the first study on the isolation and characterization of B. thuringiensis from olive-related habitats in Turkey. No correlation was observed between the cry genotypes and insecticidal crystal shapes of the isolates. Restriction profiles of 23% of the isolates were found to be different from those of the 10 reference strains used.State Planning Organization, DPT (2002K-120390

Sperm parametrelerinin ICSI başarısındaki rolü

Introduction: In this study, we aimed to investigate the relation between the sperm parameters and fertilization and pregnancy rates in intracytoplasmic sperm injection (ICSI) method which is applied to infertile couples. Materials and Methods: Four hundred thirty six couples who applied to Ege University Family Planning and Infertility Research and Treatment Center between January 2000 and January 2002 period were enrolled to the study. Infertile couples of males were evaluated by physical examination, spermiogram and hormone profile. Sperm morphologies were evaluated with Kruger criteria. Results: One hundred twenty two of 493 infertile males (24.74%) had oligozoospermia, 59 of them had asthenozoospermia (11.96%), 67 of them (13.61%) had teratozoospermia and 188 of them oligoasthenozoospermia (38.13%). For the statistical analysis, 57 (11.56%) infertile patients with normal sperm parameters were considered as the control group. The fertilization rates were as follows: 70.9% in oligozoospermic group, 72.8% asthenozoospermic group, 67.4% in teratozoospermic group, 66.7% oligoasthenozoospermic group and 79% for the control group. (p=0.27) Pregnancy rates in oligozoospermic group was 41.3%, in asthenozoospermic group was 43.9%,in teratozoospermic group 35.9%, in oligoasthenozoospermic group was 46.4% and for the control group was 49.1% (p=0.632). Conclusion: We did not see any difference between the disorders of sperm parameters oligozoospermia, asthenozoospermia, teratozoospermia, oligoasthenozoospermia. ICSI seems to be a successful method in the treatment of male infertility.Bu çalışmamızda, infertil çiftlere uygulanan ICSI yönteminde sperm parametreleri ile fertilizasyon ve hamilelik oranları arasındaki ilişkiyi saptamayı araştırdık. Ocak 2000-Ocak 2002 tarihleri arasında Ege Üniversitesi Aile Planlaması, Kısırlık Uygulama ve Araştırma Merkezi’ne başvuran 493 çift değerlendirmeye alındı. İnfertil erkek hastalar fizik muayene, semen analizi ve hormon profili ile değerlendirildi. Hastalara ICSI yöntemi uygulanarak fertilizasyon ve gebelik oranları karşılaştırıldı. Yapılan değerlendirmede toplam 493 infertil erkeğin 122’sinde oligozoospermi (%24.74), 59’unda astenozoospermi (%11.96), 67’sinde teratozoospermi (%13.61) ve 188’inde oligoastenozoospermi saptandı (%38.13). Ayrıca normal sperm parametreleri olan 57 infertil hasta (%11.56) kontrol grubu olarak çalışmaya dahil edildi. Fertilizasyon oranları; oligozoospermik grupta %70.9, astenozoospermik grupta %72.8, teratozoospermik grupta 67.4, oligoastenozoospermik grupta %66.7 ve kontrol grubunda ise %79 olarak saptandı (p=0.27). Gebelik oranları incelendiğinde oligozoospermik grupta %41.3, astenozoospermik grupta %43.9, teratozoospermik grupta %35.9, oligoastenozoospermik grupta %46.4 ve kontrol grubunda ise %49.1 oranları elde edildi (p=0.632). Çalışmamızda farklı sperm parametrelerinde bozukluk (oligozoospermi, astenozoospermi, teratozoospermi, oligoastenozoospermi) olan hastalara uygulanan ICSI yönteminde, fertilizasyon ve gebelik oranları arasında istatistiksel olarak anlamlı bir fark saptanmadı. ICSI erkeğe bağlı infertilite tedavisinde başarılı bir yöntem olarak görünmektedir

The role of sperm parameters on ICSI success

Bu çalışmamızda, infertil çiftlere uygulanan ICSI yönteminde sperm parametreleri ile fertilizasyon ve hamilelik oranları arasındaki ilişkiyi saptamayı araştırdık. Ocak 2000-Ocak 2002 tarihleri arasında Ege Üniversitesi Aile Planlaması, Kısırlık Uygulama ve Araştırma Merkezi’ne başvuran 493 çift değerlendirmeye alındı. İnfertil erkek hastalar fizik muayene, semen analizi ve hormon profili ile değerlendirildi. Hastalara ICSI yöntemi uygulanarak fertilizasyon ve gebelik oranları karşılaştırıldı. Yapılan değerlendirmede toplam 493 infertil erkeğin 122’sinde oligozoospermi (%24.74), 59’unda astenozoospermi (%11.96), 67’sinde teratozoospermi (%13.61) ve 188’inde oligoastenozoospermi saptandı (%38.13). Ayrıca normal sperm parametreleri olan 57 infertil hasta (%11.56) kontrol grubu olarak çalışmaya dahil edildi. Fertilizasyon oranları; oligozoospermik grupta %70.9, astenozoospermik grupta %72.8, teratozoospermik grupta 67.4, oligoastenozoospermik grupta %66.7 ve kontrol grubunda ise %79 olarak saptandı (p=0.27). Gebelik oranları incelendiğinde oligozoospermik grupta %41.3, astenozoospermik grupta %43.9, teratozoospermik grupta %35.9, oligoastenozoospermik grupta %46.4 ve kontrol grubunda ise %49.1 oranları elde edildi (p=0.632). Çalışmamızda farklı sperm parametrelerinde bozukluk (oligozoospermi, astenozoospermi, teratozoospermi, oligoastenozoospermi) olan hastalara uygulanan ICSI yönteminde, fertilizasyon ve gebelik oranları arasında istatistiksel olarak anlamlı bir fark saptanmadı. ICSI erkeğe bağlı infertilite tedavisinde başarılı bir yöntem olarak görünmektedir.Introduction: In this study, we aimed to investigate the relation between the sperm parameters and fertilization and pregnancy rates in intracytoplasmic sperm injection (ICSI) method which is applied to infertile couples. Materials and Methods: Four hundred thirty six couples who applied to Ege University Family Planning and Infertility Research and Treatment Center between January 2000 and January 2002 period were enrolled to the study. Infertile couples of males were evaluated by physical examination, spermiogram and hormone profile. Sperm morphologies were evaluated with Kruger criteria. Results: One hundred twenty two of 493 infertile males (24.74%) had oligozoospermia, 59 of them had asthenozoospermia (11.96%), 67 of them (13.61%) had teratozoospermia and 188 of them oligoasthenozoospermia (38.13%). For the statistical analysis, 57 (11.56%) infertile patients with normal sperm parameters were considered as the control group. The fertilization rates were as follows: 70.9% in oligozoospermic group, 72.8% asthenozoospermic group, 67.4% in teratozoospermic group, 66.7% oligoasthenozoospermic group and 79% for the control group. (p=0.27) Pregnancy rates in oligozoospermic group was 41.3%, in asthenozoospermic group was 43.9%,in teratozoospermic group 35.9%, in oligoasthenozoospermic group was 46.4% and for the control group was 49.1% (p=0.632). Conclusion: We did not see any difference between the disorders of sperm parameters oligozoospermia, asthenozoospermia, teratozoospermia, oligoasthenozoospermia. ICSI seems to be a successful method in the treatment of male infertility