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Antibiyotikler, bakteriyel infeksiyonları önlemek için temel tedavi edici seçenektir. Ancak, bu konuda bakteriyel direncin artmasından dolayı dezavantaj da vardır. Bu yüzden bitkiler üzerine yapılan antimikrobiyal çalışmalar önemli bir alternatif olmuştur. Bu çalışmada, 24 hastadan alınan kültür örneklerinden izole edilen mikroorganizmalara karşı Sinop’tan toplanan Jurinea kilae, Isatis arenaria, Verbascum degenii ve Pancratium maritimum bitki türlerinden farklı çözücüler (metanol, etanol ve su) kullanılarak elde edilen ekstraktların antimikrobiyal aktivitelerinin araştırılması amaçlanmıştır. Klinik örneklerden izole edilen mikroorganizmaların cins ve tür düzeyinde tanımlamaları VITEK 2 Compact (bioMérieux, Fransa) sisteminde yapılmıştır. Bitkiler, Sinop ilinde farklı noktalardan toplanmış ve tür teşhisi yapıldıktan sonra diğer işlemler için saklanmıştır. Bitki örneklerinden ekstraksiyon için metanol, etanol ve su kullanılmıştır. Antimikrobiyal aktivite, disk difüzyon ve mikrodilüsyon yöntemleri kullanılarak belirlenmiştir. Disk difüzyon sonuçlarına göre, özellikle I. arenaria ve V. degenii türlerinden elde edilen metanol ve etanol ekstraktları Klebsiella pneumoniae ssp. ozaenae, Proteus, Serratia, Staphylococcus, Kocuria ve Candida’ya karşı daha yüksek aktivite (10-26±1.4 mm) göstermiştir. Ayrıca ekstraktların, Gram (-) bakteriler ve mayalara nazaran Gram (+) bakterilere karşı daha etkili oldukları saptanmıştır. Aktif ekstraktların minimum inhibisyon konsantrasyonu (MİK) değerleri farklı organizmalara karşı 250-1000 µg mL-1 aralığında bulunmuştur. Bu çalışmanın sonuçları, I. arenaria ve V. degenii türlerinden elde edilen metanol ve etanol ekstraktlarının alternatif doğal antimikrobiyal ajanlara sahip olduğunu vurgulamaktadır. Böylece, bu sonuçlara göre I. arenaria ve V. degenii metanol ve etanol ekstraktlarının antimikrobiyal bileşikler içerdiği ve bunun yeni ilaç geliştirilmesinde kullanılabileceği önerilebilir. Bu çalışma aynı zamanda klinik kökenli infeksiyonlarla ilgili halk sağlığı risklerinin azalmasına önemli bir katkı sağlayabilir.

Amaç: Bu çalışmada, Sinop İlindeki balıkçılardan temin edilen hamsi ve zargana balık örneklerinden izole edilen Vibrio türlerinin karekterizasyonu ile bu izolatların antibiyotik dirençlilikleri, plazmid DNA ve SDS-PAGE (sodyum dodesil sülfat poliakrilamid jel elektroforezis) toplam hücre protein profillerinin belirlenmesi amaçlanmıştır. Yöntem: Balık örnekleri taze olarak alınmış, buz üzerinde korunarak laboratuvara getirilmiş ve hemen analize alınmıştır. Balık örnekleri 25 g tartılarak zenginleştirme amacıyla %3 NaCl içeren steril alkali peptonlu su (225 mL) besiyerine ilave edilmiş ve 37 OC'de 18-24 saat inkübasyona bırakılmıştır. İnkübasyon sonrası Tiyosülfat Sitrat Safra Sükroz (TCBS) agar besiyerinde gelişen sarı, yeşil veya mavi-yeşil koloniler Tripton Soya Agar (TSA) besiyerine ekilmiş ve izolatların daha sonraki tanımlaması için bir gece süreyle inkübe edilmiştir. İzolatlar morfolojik, fizyolojik ve biyokimyasal testlere göre tanımlanmıştır. Bunlara ek olarak izolatların yedi farklı antibiyotiğe karşı dirençliliği disk-difüzyon yöntemiyle belirlenmiştir. Ayrıca, izolatların SDSPAGE hücresel protein ve plazmid DNA profilleri de çıkartılmıştır. Bulgular: Toplam 44 adet balık örneği Vibrio türlerinin belirlenmesine yönelik çalışılmış ve morfolojik, biyokimyasal özellikleri yönünden incelenmiştir. İnceleme sonucunda; 12 adet Vibrio spp. belirlenmiştir. zolatların, seftazidime %66,6, gentamisine %25, tetrasikline %16,6, seftriazona %91,6, amikasine %16,6, ofloksine %25 ve penisilin G'ye %58,3 oranında antibiyotik dirençlikleri saptanmıştır. Ayrıca, test edilen izolatların MARI (Çoklu Antibiyotik Dirençlilik İndeks) değeri 0,41 oranında bulunmuştur. Vibrio spp. izolatlarının SDSPAGE toplam hücre protein profillerine göre oluşturulan nümerik analiz %80 ve üzerinde benzerlik seviyesinde iki temel küme oluşturmuştur. Vibrio spp. izolatların tümünde 1-4 plazmid DNA (1590 - 27000 bp aralığında) tespit edilmiştir. Sonuç: Sonuç olarak, balıklar yakalandıktan sonra yeterince dondurularak, pişirilerek ve uygun şartlarda depolanarak; ayrıca paketleme esnasında çapraz bulaşlara engel olunarak mikrobiyal enfeksiyonların önlenebileceğini söylemek mümkündür. Ayrıca, bakterilerden dolayı kıyı sularında meydana gelen bulaşlar, deniz suyu kaynaklarının kalitesinde düşüşlere ve bu yolla da insan sağlığında riske ve ekonomik kayıplara yol açmaktadır. Çalışmamız, sağlık için potansiyel risk olabilen ve balıklardan izole edilebilen, antibiyotiklere dirençli Vibrio spp. suşlarının varlığını göstermektir, Objective: In this study it was aimed to characterize the Vibrio species isolated from anchovy and garfish samples obtained from fishermen and to determine the antibiotic resistances, plasmide DNA and SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis) whole cell protein profiles of these isolates. Method: The fish samples were taken as fresh, kept in ice during transport to the laboratory and immediately taken for analysis. The fish samples weighed as 25 grams were added into 225 mL of sterile alkaline peptone water supplemented with 3% NaCl broth for enrichment purpose and incubated at 37 OC for 18 to 24 hours. After incubation, yellow, green or blue-green colonies growing on Thiosulphate Citrate Bile Salt Sucrose (TCBS) agar were subcultured on Triptone Soya Agar (TSA) and incubated overnight for further identification of the isolates. The isolates were identified according to morphological, physiological and biochemical tests. Moreover, the resistances against seven different antibiotics of isolates were determined by disk-diffusion method. In addition, the SDS-PAGE cell protein and plazmid DNA profiles of isolates were determined. Results: Totallly 44 fish samples were tested for determining Vibrio species and were analyzed according to their morphological, biochemical properties. At the end of the analysis; 12 Vibrio species were determined. The isolates were resistant against to ceftazidime 66.6%, gentamisine 25%, tetracycline 16.6%, ceftriaxone 91.6%, amikacine 16.6%, ofloxacine 25% and penicillin G 58.3%. In addition, The MARI (Multiple Antibiotic Resistance Index) values of tested strains were found at the rate of 0.41. The numerical analysis of SDS-PAGE whole-cell protein profiles of Vibrio spp. isolates revealed two major clusters at similarity levels of 80% and above. The 1-4 plasmide (range 1590 - 27000 bp) in all of the isolates were determined. Conclusion: As a result, it is possible to say that microbial infections can be prevented by adequate freezing and cooking, proper storage and processing after harvesting and avoidance of cross-contamination during fish handling. In addition, the coastal contamination due to bacteria leads to decline in the quality of seawater resources and so leads to risks to human health and economic losses. Furthermore, our study indicated that the presence of antibiotic resistant Vibrio spp. strains isolated from fishes might be potential risk for health.

Five reference Bacillus species and 17 native alkaliphilic Bacillus strains, isolated from the water of Lake Van and the soil surrounding it, were identified using phenotypic characteristics and extracellular protein profiles by the method of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). According to the phenotypic characteristics, all the native isolates were Gram-positive, aerobic, endospore-forming, motile, facultatively alkaliphilic Bacillus spp. It was also found that the native alkaliphilic Bacillus isolates were distinguished phenotypically from other Bacillus species. A numerical analysis based on the resulting extracellular protein profiles revealed 5 distinct clusters: native isolates and B. megaterium formed 4 clusters that varied between 69% and 100% similarity, and the other reference Bacillus species formed a cluster with a similarity of above 70%. The research showed that extracellular protein profiles obtained by SDS-PAGE provide an effective approach to the investigation of taxonomic relationships within native alkaliphilic Bacillus isolates and other Bacillus species.

The effects of 9 commercial pesticides used commonly in agriculture on spore germination and larvicidal activity of the mosquito pathogenic B. thuringiensis var. israelensis and B. sphaericus 2362 strain were examined. Six of the pesticides was had an inverse impact on spore germination and growth of the test strains. It was determined that the pesticides with the most adverse effect were copper sulfate, methiocarb, and dalapon on spore germination of the microorganisms. Higher concentrations or MIC values of the pesticides reduced the heat-resistant spore numbers of both bioinsecticides, suppressing spore germination. These results indicated that the spores of the bacteria are sensitive to chemical pesticides at the same level. Moreover, the study revealed that the spore germination and larvicidal activity of both biological control agents are affected by pesticides equally. The findings should be considered for assessing the effects of pesticides when both microbial agents are used in field applications.

In the present study nine reference Bacillus strains were characterized by whole-cell protein profiles using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A numerical classification of the protein profiles revealed two distinct clusters at 47% similarity level. Cluster 1 comprised four strains belongs to B. subtilis and B. megaterium species at similarity levels changed between 67 and 85%, while cluster 2 consisted five strains belongs to various Bacillus species at similarity levels changed between 50 and 70%. The strains of the cluster 2 were clearly separated from that of cluster 1 by numerical analysis. Our results indicates that SDS-PAGE method combined with computerized analysis of cellular protein profiles provide an effective approach to investigate of taxonomic relationships within Bacillus species., Bu çalışmada, dokuz referans Bacillus ırkı sodyum dodesil sülfat poliakrilamid jel elektroforezi kullanılarak elde edilen toplam hücresel protein profillerine göre karakterize edildi. Protein profil sonuçları esas alınarak yapılan nümerik sınıflandırma %47 benzerlik düzeyinde iki ayrı grup (cluster) oluşturdu. Grup 1 benzerlik düzeyi %67-85 arasında değişen B. subtilis ve B. megaterium türlerine ait dört ırkı; grup 2 benzerlik seviyesi % 50-70 arasında değişen farklı Bacillus türlerine ait beş ırkı içermektedir. Nümerik analizde grup 1’e ait ırklar grup 2’den açıkça ayrılmaktaydı. Bizim sonuçlarımız, SDS-PAGE yöntemiyle elde edilen hücresel protein profillerinin bilgisayar analizleriyle birleştirilmesinin Bacillus türlerinin taksonomik ilişkilerinin incelenmesinde etkili bir yaklaşım sağladığını gösterdi.

In the study, the changes in the serum, liver, kidney protein and genomic DNA profiles in Wistar albino rats administrated to 7,12-dimethylbenz(a)anthracene (DMBA) and Plantagomajor L. were investigated. SDS-PAGE profiling of the serum proteins showed that the levels of two proteins molecular weighing 140.8 kDa and 46.8 kDa were significantly lower on the 0th day of treatment than on the 60th day of application in the groups. Furthermore, two proteins (34.5 and 22.3 kDa) were solely present in DMBA-treated liver cell lysates. On the contrary, the kidney protein profiles did not show significant banding variations in groups. Genomic DNA analysis also confirmed that DNA extracted from liver and kidney cells did not fragment in any of the groups after the 60th study day. The proteins 34.5 and 22.3 kDa in liver cell lysates at the 60th day in DMBA-treated group could be extra-prognostic indicators for severe DMBA toxicity.

A new and simple high-performance liquid chromatography method was developed and validated for the simultaneous determination of retinol, retinyl palmitate, beta-carotene, alpha-tocopherol and vitamin C in rat serum treated with Plantago Major L. and 7,12 dimethylbenz[a] anthracene. High-performance liquid chromatography analysis was performed utilizing an Inertsil ODS3 reversed phase column with methanol-tetrahydrofuran-water as mobile phase under gradient conditions, at 1.5 mL min(-1) flow rate and 25 degrees C. Diode-array detection was at 325, 450, 290 and 270 nm (retinol and retinyl palmitate), beta-carotene, alpha-tocopherol and vitamin C, respectively and runnig time 18 min. The high-performance liquid chromatography assay and extraction procedure proposed are simple, rapid, sensitive and accurate. The method was then applied for the determination of retinol, retinyl palmitate, beta-carotene, alpha-tocopherol and vitamin C in rat serum. Results of this study demonstrated that; at 60th day DMBA-treated group, there was a significant decrease in vitamin levels compared to the levels of control group. A significant increase was observed in vitamin levels of 7,12 dimethylbenz[a]anthracene+Plantago Major L.-treated group compared to the DMBA-treated group. Additionally, the results obtained in the study are found to be in agreement with data reported in the literature.

Present study has been conducted for finding out the total protein profile of bacterial strain Streptomyces sps by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Total 139 isolates of Streptomyces have been isolated from the soil. Amongst all isolated strain, total 20 isolates were used for getting protein profile by SDS PAGE. Amongst all isolates, 20 isolates were selected for protein profiling and these were divided in two groups. Two strains of Streptomyces i.e. S. violaceus and S. albidoflavus were selected as a reference strain for both groups. Band profile were analyzed and assessed by computer added program BioRad Quantity with the use of Unweighted Pair Group Method of Analysis (UPGMA). As a result of this computer assisted numeric analysis study, approximately 40 different types of protein bands were reported between 10 or 100 kD molecular weight. Analysis of acquired dendogram on the basis of similarities ratios, all 40 proteins can be divided in to 7 groups. In addition, the isolates A4B3G, D145B, S5036.6 and reference isolate S. violaceus were available in the same group, while 805A, C804B, F1705 isolates and reference sample S. albidoflavus were detected in the same group. The test organisms which were similar to each other in terms of morphological and biochemical characters delivered the same protein bands. SDS-PAGE method is an effective method in terms of determining taxonomical relations between the various species of genus Streptomyces.

In the present research, a total 28 bean cultivars grown in Turkey were characterized by the numerical analysis of seed protein profiles obtained by using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) technique. The numerical analysis of seed protein profiles generated two major clusters at similarity levels of 82.52% or above. The first cluster included 5 reference bean cultivars at the intra-cluster similarity values of 91.25% or above. The second cluster comprised the all remaining genotypes at similarity values between 84.46% and 100%. Our results also indicated that Gina standard cultivar together with all eastern genotypes belonged to subcluster IIc, having the highest intra-cluster similarities between 95.54% and 100%. In this sense, it is suggested that probably there may be a very close genetic relationship between eastern genotypes and Gina cultivar. In conclusion, this study indicated that the numerical analysis of seed protein profiles offer no final answer to the identity of bean genotypes, however, the method can provide of extra banding pattern for the discrimination of bean genotypes cultivated in Turkey.

In the study, a total of 41 strains of Staphylococcus aureus including 38 clinical isolates and 3 reference strains were characterized according to biotyping, plasmid profiles and the numerical analysis of the protein profiles. The antibiogram results showed that the isolates were resistant against the tested antibiotics, except for vancomycin. Furthermore, plasmid profile analysis confirmed that the strains consisted of only one plasmid. Present findings indicated that the whole-cell and extracellular protein profiles obtained by using SDS-PAGE methods to be good typing tool for the differentiation of S. aureus strains at the species level, not strain level. However, Native-PAGE of whole-cell proteins was the most reliable and rapid method for differentiation between MRSA and ordinary S. aureus strains comparing to other applying PAGE techniques. In addition, it was determined that the same clone was responsible for most cases of MRSA and other S. aureus epidermic among surveyed hospitals. In conclusion, this study supposed that together application at least two different polyacrylamide gel electrophoresis (PAGE) techniques can be offer accurate and an effective approach to the investigation of taxonomic relationships within human pathogenic S. aureus strains.

In the present study, the antimicrobial activities of 10 different plant species growing in Van region were determined against 5 Gram-positive (Staphylococcus aureus ATCC 25923, S. epidermidis, Micrococcus iuteus ATCC 9345, Bacillus cereus ATCC 7064 and B. thuringiensis var. israelensis 2 Gram-negative (Escherichia coli ATCC 4230 and Enterococcus faecalis ATCC 29212) and 3 yeast (Candida krusei ATCC 6258, C. parapffosis ATCC 22019 and C. albicans ATCC 14053) $trains by using disc diffusion method. White the extracts of Saivia kronenburgii, Antriseus nenorosa and Cephaloria setosa were the highest antimicrobial activity against all tested strains, it was found that the extracts of Thymus transcaucasjcusj Morus albay Pinus nigra subsp. pallasiana and Hypericum perforatum had no antimicrobial activity. However, the extracts of Syrjngae vulgaris and Salix alba exhibited Iow antimicrobial activity against tested microorganisms. Also the extract of Alyssum meniocoides was most effective against yeast strains (1216 mm). Consequently, the research indicated that Tris-HCI extracts of four plants extract that having good antibacterial and antifungal activity can be used as natural antimicrobial agents., Bu çalışmada, Van'da yetişen 10 farktı bitki türünden elde edilen Tris-HCI özütlerinin antimikrobiyal aktiviteleri 5 Gram-pozitif (Staphylococcus aureus ATCC 25923, S. epidermidis, Micrococcus luteus ATCC 9345, Bacillus cereus ATCC 7064 ve B, thuringjensis var. israelensis), 2 Gram-negatif (Escherichja coli ATCC 4230 ve Enterococcus faecalis ATCC 29212) ve 3 maya (Candida krusei ATCC 6258, C, parapilosis ATCC 22019 ve C. albicans ATCC 14053) türüne karşı disk difüzyon yöntemi kullanılarak belirtendi. Salvia kronenburgii, Antrjseus nenorosa ve Cephaloria setosa özütlerinin test edilen bütün suşlara karşı en yüksek antimikrobiyal etkiye sahip olmalarına karşın, Thymus franscaucasicus, Morus alba, Pinus nigra subsp. pallasiana ve Hypericum perforatum özütlerinin etkisiz oldukları bulundu. Diğer taraftan, Syringae vulgaris ile Salix alba özütleri düşük antimikrobiyal aktivite gösterdi. Ayrıca, Alyssum meniocoides özütünün funguslara (12-16 mm) daha etkili olduğu belirlendi. Sonuç olarak, bu araştırma antibakteriyal ve antifungal etkiye sahip olduğu belirlenen 4 bitki türünün Tris-HCl özütlerinin doğal antimikrobjyal ajan olarak kullanılabileceğini göstermektedir.

In the present study, the effect of Nettle (Urtica dioca L.) on trace elements and serum protein patterns in female rabbits administrated to 7,12-Dimethylbenz(a)anthracene (DMBA) was investigated. Rabbits were divided into equal four groups. Group I was control group (n = 8), group 11 (n = 8), treated with DMBA (1 mg/kg.d), group III treated with DMBA (I mg/kg.d) and methanol extract of U. dioica 35 mg/kg/d and group IV (n = 8) treated with DMBA (1 mg/kg.d) and aqueous extract of U. dioica 35 mg/kg/d. The results show that on 60th day DMBA-treated group were compared with the control group levels, a significant decrease (p < 0.05) was observed in the Zn. DMBA and methanol extract of U. dioica treated group were compared with the control group levels, a significant increase (p < 0.05) was observed in the Cu. Serum Fe and Mn levels among groups were determined not to be statistically significant (p > 0.05). It was found that U. dioica not affected on trace elements level and serum protein patterns in rabbits subjected to DMBA.

In the present study, plasmid DNA profiles and susceptibility to various antibiotics of a total 35 methicillin resistant Staphylococcus aureus (MRSA) strains isolated from clinical specimens were investigated. All MRSA strains were susceplible to varçomycin, but it was determined they were resistant to beta-lactams, quinolones, aminogliycosides, macrolid, ghloramphenicol, piperacillin + tazobactam and trimetroprim + sulfamethoxazole. The agarose gel electrophoresis reşults çonfirmed that all strains consisted of only one plasmid, molecular weight 13.868 bp, The results of this research indicated that in ihe effective therapy of infections caused by S. aureuş strains resistant to methicillin and multiple antibiotics, glycopeptide antibiotics should be chosen., Bu çalışmada, klinik örneklerden izole edilen 35 adet metisiline dirençli Staphylococcus aureus (MRSA) suşunun plazmit DNA profilleri ve farklı antibiyotiklere karşı duyarlılıkları incelenmiştir. Bütün MRSA suşlarının vankomisine duyarlı, fakat beta-laktamlar, kinolonlar, aminoglikozitter, makroliti, kloramfenikol, piperasilin + lazobaktam ve trimetroprim + sülfametoksazole dirençli oldukları belirlenmiştir. Agaroz jel elektroforez sonuçları bütün suşların 13.868 bp molekül ağırlığında tek bir plazmit içerdiğini ortaya koymuştur. Bu çalışmanın sonuçları, metisiline ve birçok antibiyotiğe dirençli S. aureus suşlarının neden olduğu infeksiyonların etkin tedavisinde glikopeptit grubu antibiyotiklerin tercih edilmesi gerektiğini göstermiştir.

In the present study, 21 Candida albicans strains were investigated using the commercial kit API 20C AUX system and the numerical analysis of whole-cell protein profiles. The results of the commercial kit confirmed that the all the strains belonged to C. albicans species. However, the research indicated that SDS-PAGE of polypeptides of whole-cell extracts can provide more valuable taxonomic information than conventional yeast test kits at the subspecies level. Despite the fact that C. albicans subtypes isolated from different anatomical sites had similar protein profiles, there were some distinctive protein bands. Numerical analysis of whole-cell protein profiles of all strains revealed 2 major clusters at similarity degrees of between 46.26 and 100%. Moreover, the results of numerical analysis confirmed that each cluster had characteristic and distinctive protein profiles. The research showed that, the morphological examination of yeast isolates remains essential to obtaining a correct identification, both the commercial yeast kit system and the numerical analysis of whole-cell protein patterns can be useful for the more reliable identification of C. albicans strains.

In the present study, 13 E. coli strains were characterized using phenotypic properties, plasmid and protein profiles. According to agarose gel electrophoresis analysis it was confirmed that all strains consisted of at least one plasmid. A numerical analysis based on whole-cell protein patterns revealed 2 distinct major groups with a similarity level of above 71 %. Afterwards, a numerical analysis based on extracellular protein patterns revealed 3 difference major groups with a similarity level of above 60 %. The research indicated that whole-cell and extracellular protein profiles obtained by SDS-PAGE did not provide an effective approach to the investigation of taxonomic relationship among the tested E. coli strains. However, it determined that extracellular protein patterns were more difference than whole-cell protein profiles., Bu çalışmada, 13 adet E. coli suşu fenotipik özellikleri, plazmit ve protein profillerine göre karakterize edildi. Agaroz jel elektroforez analiz sonuçlarına göre bütün suşların en az bir adet plazmit bulundurduğu belirlendi. Toplam hücre protein profil sonuçları esas alınarak yapılan nümerik analiz, benzerlik düzeyi % 71'in üzerinde 2 ayrı büyük grup ortaya koydu. Ayrıca, hücre dışı protein profil sonuçları esas alınarak yapılan nümerik analiz ise benzerlik düzeyi % 60'ın üzerinde 3 ayrı büyük grup ortaya koydu. Bu araştırma, SDS-PAGE yöntemiyle elde edilen toplam hücre ve hücre dışı protein profillerinin test edilen E. coli suşlarının taksonomik ilişkilerinin incelenmesinde etkili bir yaklaşım sağlamadığını gösterdi. Bununla birlikte, hücre dışı protein profillerinin toplam hücre protein profillerine göre daha iyi ayırım sağladığı da belirlendi.

In the present study, seven reference Bacillus species and a total of eighteen new facultative alkaliphilic Bacil-lus strains, isolated from the water of Lake Van and its soil surroundings, were identified using phenotypic charac-teristics and the numerical analysis of whole-cell protein profiles. According to morphological, biochemical and physiological characteristics, it was found that the new isolated strains belonged to Bacillus genus. In addition, the research indicated that SDS-PAGE of polypeptides of whole-cell extracts can provide more valuable taxonomic information than conventional tests based on the pheno-typic characteristics at the species and subspecies levels. Numerical analysis of whole-cell protein profiles of test strains revealed 4 basic clusters (I-IV) at dissimilarity val-ues of 18.9 % or above. The results of numerical analysis confirmed that each cluster had characteristic and distinc-tive protein profiles. Indeed, this study showed that the application of numerical analysis, coupled with the utiliza-tion of a standardized identification system instead of sim-ple quantitative comparison of protein patterns, greatly enhanced the utilization of whole-cell protein profiles for identification of the facultative alkaliphilic Bacillus species.

Five reference Bacillus species and 17 native alkaliphilic Bacillus strains, isolated from the water of Lake Van and the soil surrounding it, were identified using phenotypic characteristics and extracellular protein profiles by the method of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). According to the phenotypic characteristics, all the native isolates were Gram-positive, aerobic, endospore-forming, motile, facultatively alkaliphilic Bacillus spp. It was also found that the native alkaliphilic Bacillus isolates were distinguished phenotypically from other Bacillus species. A numerical analysis based on the resulting extracellular protein profiles revealed 5 distinct clusters: native isolates and B. megaterium formed 4 clusters that varied between 69% and 100% similarity, and the other reference Bacillus species formed a cluster with a similarity of above 70%. The research showed that extracellular protein profiles obtained by SDS-PAGE provide an effective approach to the investigation of taxonomic relationships within native alkaliphilic Bacillus isolates and other Bacillus species., Bu çalışmada, 5 referans Bacillus türü ve Van Gölü suyu ve çevresindeki topraklardan izole edilen toplam 17 yerel alkalofilik Bacillus izolatı fenotipik özellikleri ve sodyum dodesil sülfat poliakrilamid jel elektroforez kullanılarak elde edilen ekstraselülar protein profillerine göre identifiye edilmiştir. Fenotipik özelliklerine göre, yerel izolatların tümünün Gram pozitif, aerobik, endospor oluşturan, hareketli ve fakültatif alkalofilik Bacillus cinsine ait türler olduğu belirlenmiştir. Ayrıca, yerel alkalofilik izolatların fenotipik olarak diğer referans Bacillus türlerinden farklı olduğu bulunmuştur. Ekstraselülar protein profil sonuçları esas alınarak yapılan nümerik analiz; yerel izolatlar ve B. megaterium'u içeren benzerlik seviyesi % 69 ve % 100 arasında değişen dört grup ve benzerlik seviyesi % 70'in üzerinde diğer referans Bacillus türlerini içeren bir grup olmak üzere beş ayrı grup ortaya koymuştur. Bu araştırma, SDS-PAGE yöntemiyle elde edilen ekstraselülar protein profillerinin yerel alkalofilik Bacillus izolatları ve diğer Bacillus türlerinin taksonomik ilişkilerinin incelenmesinde etkili bir yaklaşım sağladığını göstermiştir.

In this study, 12 different commercial pesticides, in-cluding 5 fungicides, 5 herbicides, 1 aphicide and 1 acari-cide, were investigated for their effects on spore viability, toxin stability and larvicidal activity of a mosquito patho-genic B. sphaericus 2362 strain. Minimal inhibitory con-centrations (MICs) of each pesticide were determined after 48 h incubation of B. sphaericus 2362 strain. Ac-cording to MIC results, all pesticides showed various inhibitory effects on the growth of the test bacterium. Particularly, the pesticides which contain TCA, hymexa-zol, copper sulphate, hexythiazox, 2,4-D Amine, copper inhibited spore germination significantly and bacterial growth even at the lowest tested concentrations. Our re-sults indicated that various commercial pesticides in dif-ferent concentrations reduced the spore viability signifi-cantly and heat-resistant spore numbers of B. sphaericus 2362 strain. In general, the pesticides decreased the spore numbers 104-105-fold compared to initial ones. Further-more, it was observed that the larvicidal activity was completely lost when it was grown at the concentration of MIC values or higher. Indeed, SDS-PAGE analysis con-firmed that the binary toxin proteins, 41.9 kDa and 51.4 kDa, were lost at the MIC values and higher concentra-tions although parasporal crystals did not disappear in the much lower pesticide concentration.

The effects of 9 commercial pesticides used commonly in agriculture on spore germination and larvicidal activity of the mosquito pathogenic B. thuringiensis var. israelensis and B. sphaericus 2362 strain were examined. Six of the pesticides was had an inverse impact on spore germination and growth of the test strains. It was determined that the pesticides with the most adverse effect were copper sulfate, methiocarb, and dalapon on spore germination of the microorganisms. Higher concentrations or MIC values of the pesticides reduced the heat-resistant spore numbers of both bioinsecticides, suppressing spore germination. These results indicated that the spores of the bacteria are sensitive to chemical pesticides at the same level. Moreover, the study revealed that the spore germination and larvicidal activity of both biological control agents are affected by pesticides equally. The findings should be considered for assessing the effects of pesticides when both microbial agents are used in field applications., Bu çalışmada, zirai amaçla yaygın olarak kullanılan dokuz ticari pestisitin sivrisinek patojenleri B. thuringiensis var. israelensis ve B. sphaericus 2362 ırklarının spor çimlenmesi ve larvasidal aktivitesi üzerine etkileri incelenmiştir. Üç tanesi hariç, bütün pestisitler test ırklarının büyümesi ve spor çimlenmesini olumsuz etkilenmiştir. Mikroorganizmaların spor çimlenmesi üzerine olumsuz etki gösteren pestisitlerin bakır sülfat, methiocarb ve dalapon olduğu belirlenmiştir. Pestisitlerin MÜK veya daha yüksek konsantrasyonları spor çimlenmesini baskılayarak her iki biyoinsektisitin ısıya dayanıklı spor sayılarınn düşürmüştür. Bu sonuçlar bakteri sporlarrının kimyasal pestisitlere karşı aynı seviyede duyarlı olduğunu göstermiştir. Ayrıca, çalışmada pestisitlerle muamele edilmiş biyoinsektisitlerin larvasidal aktivitelerinin de benzer seviyede etkilendiği belirlenmiştir. Her iki mikrobiyal kontrol ajan arazi uygulamalarında kullanğında pestisitlerin etkilerinin tespit edilmesine ait bulgular dikkate alınmalıdır.

Spore viability and larvacidal activity of B. sphaeri-cus 2362 and 1593 strains were monitored in media con-taining Gramaxone® herbicide. Minimal Inhibitory Con-centrations (MICs) of the herbicide were determined for each of the mosquito pathogenic strains. Addition of paraquat into the media suppressed the spore germination after 24 and 48 h, but no influence was observed on parasporal inclusions containing insecticidal toxin pro-teins. SDS-PAGE analysis confirmed the presence of the so-called binary toxin comprising two proteins that are deposited in parasporal crystals treated with the herbicide. Moreover, the number of heat-resistant spores and total cell numbers of test strains were reduced drastically when test strains were grown in media containing 50, 100 and 200 mg ml-1 of paraquat. In contrast, there were no differ-ences between heat-resistant spore counts of both strains when grown under diet concentrations of less than 50 mg ml-1. This study revealed that B. sphaericus 2362 strain has a higher mosquito larvacidal activity than 1593 strain in the presence of paraquat in the media. Thus, the use of Gramaxone® herbicide can have harmful effects on mos-quitocidal biological control agents.

6. ÖZET Bu çalışmada, altısı fungusit, beşi herbisit ve biri afısit ve biri de akarisit olmak üzere toplam 13 adet ticari pestisitin sivrisinek patojeni olan H. sphaericus 2362 susunun gelişmesi, canlılığı, sporulasyonu, toksin stabilitesi ve larvasidal aktivitesi üzerine olan etkileri incelendi. Araştırmada, her pestisit için bakteri gelişiminin engellendiği en düşük pestisit konsantrasyonları (MİK) belirlendi. MİK denemelerinde; Tachigaren 70 WP pestisitinin 3,281 (mg/ml) hymexazol içeren konsantrasyonunda, Nissorun 5 EC pestisitinin 1,250 (mg/ml) hexythiazox içeren konsantrasyonunda, Agro D-Amin ile Cornox Amin pestisitlerinin 12,5 ve 62,5 (mg/ml) 2,4-D amin içeren konsantrasyonlarında, Nata Granül pestisitinin 1,187 (mg/ml) TC A içeren konsantrasyonunda, Koruma Göztaşı pestisitinin 1,530 (mg/ml) bakır sülfat içeren konsantrasyonunda ve Bakır WP ile Çaptan 50 WP Stauffer pestisitlerinin 3,90 (mg/ml) bakır ve çaptan içeren konsantrasyonlarında bakteri gelişiminin engellendiği bulundu. Ayrıca Primor 50 WG, Tordon 101 Mixture, Tefralin EC, Kimyagerler Pensikol ve Anvil pestisitlerinin farklı miktarda etken madde içeren bütün konsantrasyonlarında ise bakterinin normal olarak gelişip sporlanabildiği tesbit edildi. Buna ilaveten, belirlenen minimal inhibitor konsantrasyonları (MİK) ve daha yüksek pestisit konsantrasyonlarında bakterinin, 41,9 ve 51,4 kDa'luk toksin proteinlerini kaybettiği faz-kontrast mikroskobisi ve SDS-PAGE analizleriyle de doğrulandı. Bakteriyal sayım sonuçlarına göre, belirlenen minimal inhibitor konsantrasyonları (MİK) ve daha yüksek pestisit konsantrasyonlarında vejetatif hücre ve canlı spor sayısının 24, 48 ve 72 saat sonunda önemli seviyede düştüğü de tesbit edildi. Diğer taratan, pestisitlerin değişik miktarlarda etkili madde içeren bütün konsantrasyonlarında 24 ve 72 saat süresince geliştirilen B. sphaericus 2362 susunun larvasidal aktiviteleri 2-3. evre C. qinquefasciatus larvalarına karşı test edildi. Yapılan denemeler sonunda, bakteri gelişiminin engellendiği ve toksin proteinlerinin görülmediği en düşük (MİK) ve daha yüksek pestisit konsantrasyonlarında larvasidal aktivitenin97 tamamen kaybolduğu, toksin proteinlerinin görüldüğü pestisit konsantrasyonlarında ise larvasidal aktivitenin gerçekleştiği belirlendi., Sonuç olarak; Primor 50 WG, Tordon 101 Mixture, Tefralin EC, Kimyagerler Pensikol ve Anvil pestisitlerinin, sivrisinek larvalarının yaşadığı habitatlara yakın çevrelerde kimyasal mücadele amacıyla kullanılmasının önemli bir biyoinsektisit olan B. sphaerictıs 2362 susunun larvasidal aktivitesini ters yönde etkilemeyeceği, fakat Tachigaren 70 WP, Nissorun 5EC, Agro D-Amin, Koruma Göztaşı, Nata Granül, Cornox Amin, Bakır WP ve Çaptan 150 WP Stauffer pestisitlerinin kullanılmasının ise bakterinin larvasidal aktivitesini önemli seviyede düşüreceği görülmektedir. 7. SUMMARY In this study. 13 different commercial pesticides including 6 fungucides, 5 herbicides. 1 aficide and 1 acaricide were investigated for their on effects viability, sporulation. toxin stability and larvicidal activity of mosquito pathogenic of B. sphaericus 2362 strain. In research, the lowest concentrations that inhibits the bacterial growth (MICs) were determined for each pesticides. In the MIC trials; it was found the bacterial growth was inhibited with Tachigaren 70 WP pesticide (containing 3,281 mg/ml hymexazol), Nissorun 5 EC pesticide (containing 1.250 mg/ml hexythiazox), Agro D-Amin and Cornox Amin pesticides (containing 12.5 and 62.5 mg/ml 2,4-D Amin), Nata Granül pesticide (containing 1.187 mg/ml TCA). Koruma Göztaşı pesticide (containing 1,530 mg/ml copper sulphate). Bakır WP and Çaptan 50 WP Stauffer pesticides (containing 3,90 mg/ml copper and çaptan). Besides, bacteria had shown normal growth and sporulation in whole concentration of Primor 50 WG. Tordon 101 Mixture. Tefralin EC. Kimyagerler Pensikol and Anvil pesticides which are containing different amount of effective substance. In the determined minimal inhibitory concentrations (MIC`s) and higher pesticide concentrations the loss of 41.9 and 51.4 kDa toxin proteins of the bacteria was confirmed by phase-contrast microscopy and SDS-PAGE. According to bacteria count results the determined minimal inhibitory concentrations (MICs) and higher pesticide concentrations the significantly decrease in vegetative cells and viabile spore numbers after 24. 48 and 72 hours was identified. On the other hand, the larvicidal activities of B. sphaericus 2362 strain which were grown in all concentration pesticides for 24 and 72 hours, containing different amounts of effective sustance have been tested against instar 2-3. C. qiiinquefasciatns larvae. The trials showed that at the lowest pesticide concentrations in which the bacterial growth was inhibited and at higher concentrations the larvicidal activity was completely abolished and at pesticide concentrations in which the toxin proteins were the larvicidal activity was still ongoing. As a result, it was shown that, the usage of Primor 50 WG. Tordon 101 Mixture. Tefralin EC. Kimyagerler Pensikol and Anvil pesticides nearby the mopsquito larvae habitats for the purpose of chemical control will not effect inversely the important microbial insecticide B. sphaericus 2362 strain larvicidal activities, but usage of Tachigaren 70 WP. Nissorun 5 EC. Agro D-Amin. Cornox Amin. Nata Granül, Koruma Göztaşı. Bakır WP and Çaptan 50 WP Stauffer pesticides will significantly decrease the larvicidal activity of the bacteria. 133