Molecular characterization of trichophyton verrucosum strains isolated from cattle by PCR-RFLP

Dermatofitler, insan ve hayvanlarda keratin içeren dokuları infekte ederek dermatofit infeksiyonuna neden olmaktadırlar. Trichophyton verrucosum sığır dermatofitozis olgularının en yaygın etkenidir. Trichophytosis, bütün dünyada hayvancılık sektöründe önemli ekonomik kayıplara neden olması yanında zoonoz olmasıyla da insan sağlığını tehdit etmektedir. Sığırlardan genellikle T. verrucosum izole edilmektedir. Sığırlar bu etkenin doğal rezervuarıdırlar. Bu çalışmanın amacı, sığırlarda hastalığa neden olan dermatofitlerinin izolasyonu ve izole edilen T. verrucosum suşlarının Internal Transcribed Spacer (ITS) bölgelerinin PCR-RFLP ile moleküler ayrımının yapılmasıdır. Bu amaçla dermatofitozisli sığırlardan 90 adet örnek alınarak kültürleri yapıldı. Bu örneklerin kültürü sonucunda 35 (%38,8) adet T. verrucosum izole ve identifiye edildi. Bu suşların DNA izolasyonu gerçekleştirilerek ITS bölgelerin amplifikasyonu gerçekleştirildi. T. verrucosum suşlarının MvaI ve HinfI enzimleri kullanılarak yapılan Restriction Fragment Lenght Polymorphism (RFLP) analizleri sonucunda bir adet RFLP profiline rastlandı. Sonuç olarak, izole edilen T. verrucosum suşlarının PCR-RFLP sonucunda tek bir profile sahip olduğu, farklı profil örneklerinin saptanması için farklı bölgelerden hatta farklı ülkelerden suşların PCR-RFLP’lerinin yapılması gerektiği kanısına varıldı.Dermatophytes infect tissues containing keratin in humans and animals, causing dermatophytosis infection. Trichophyton verrucosum is the most common agent of bovine dermatophytosis cases. Trichophytosis causes big economic lossess throughout the world and also threatens human health by being a zoonosis. T. verrucosum is usually isolated from cattle. Cattle are the natural reservoirs of this agent. The aim of this study is to isolate disease-causing dermatophytes in cattle and to carry out molecular separation of Internal Transcribed Spacer (ITS) regions of the isolated T. verrucosum strains by PCR- Restriction Fragment Lenght Polymorphism (PCR-RFLP). For this purpose, 90 samples were taken from the cattle with dermatophytosis for cultural examination. As a result of the culture of these samples, 35 (38.8%) T. verrucosum were isolated and identified. DNA isolation of these strains was made and amplification of ITS regions was performed. It was only one RFLP profile was found according to the results of RFLP analysis of T. verrucosum strains using MvaI and HinfI enzymes. At the end of study, it was founded that the isolated T. verrucosum strains showed a single profile by PCR-RFLP analysis and PCR-RFLP was a useful tool for the molecular characterization of the strains. İt was also concluded that PCR-RFLPs of strains from different regions or even from different countries might be necessary in order to detect different profiles of the tested samples

Sığır Kıymalarında Campylobacter Türlerinin Tespitinde Mini VIDAS ve Konvansiyonel Kültür Tekniğinin Etkinliği

Bu çalışma, Bolton ve Preston brothlarda zenginleştirilmiş sığır kıymalarından Campylobacter türlerinin tespitinde, mini VİDAS otomatik immunoassay system ve konvansiyonel kültür tekniğinin etkinliklerinin değerlendirilmesi amacıyla gerçekleştirildi. Erzurum ilindeki (Türkiye) yerel marketlerden toplam 92 sığır kıyma örneği toplandı. Camylobacter türlerinin tespiti için sığır kıyması örnekleri, 42oC'de 48 h zenginleştirmeyi takiben mini VIDAS (bioMérieux, France) ve paralel olarak supplement içeren besi yerlerinde test edildi. Pozitif sonuçlar, kontrol suşları (ATCC 33559, ATCC 33560) kullanılarak selektif besiyerlerinde konvansiyonel kültür tekniği ile doğrulandı. İncelenen 92 sığır kıymasının 11 (% 11.95)'i ve 10 (%10.86)'u sırası ile mini VIDAS ve konvansiyonel kültür tekniği ile pozitif bulundu. Konvansiyonel kültür tekniğine göre Mini VIDAS'ın duyarlılığının daha yüksek olduğu ve daha doğru sonuçlar verdiği saptandı. Ayrıca, her iki broth'un zenginleştirme amacıyla etkin olarak kullanılabileceği kanaatine varıldıThis study was performed to evaluate the efficiency of the mini VIDAS automated immunoassay system and conventional culture technique to detect Campylobacter species from minced beef enriched in Bolton and Preston broths. A total of 92 minced beef samples were collected from different local markets in Erzurum province in Turkey. Minced beef samples were tested for Campylobacter spp. by the mini VIDAS (bioMérieux, France) in parallel with the culture technique using supplemented agar plates following the enrichment at 42oC for 48 h. Positive results were verified by conventional culture technique with quality control strains (ATCC 33559, ATCC 33560) on selective solid media. Out of 92 minced beef samples, 11 (11.95%) and 10 (10.86%) were positive by the mini VIDAS and culture technique respectively. The sensitivity of mini VIDAS was higher, and provided more accurate results than the conventional culture technique. In addition, both broths can be used efficiently for the enrichmen

Tulareminin Serolojik Tanısı İçin In House Enzyme- Linked Immunosorbent Assay (ELISA) Prototipi ve Mikroaglütinasyon Test (MAT) Antijeninin Geliştirilmesi

Bu çalışmada, Francisella tularensis (F. tularensis)’e karşı gelişen antikorları saptamak için bir Enzyme- Linked Immunosorbent Assay (ELI SA) prototipi ve kristal viyole boyalı bir mikroaglütinasyon test (MAT) antijeni geliştirildi. Ayrıca konfirme edici Western blot (WB) tekniği de serumlara uygulandı. Alınan sonuçlara göre MAT, ELISA ve WB testlerinin tanısal duyarlılıkları %100 olarak bulunurken, özgüllükleri sırası ile %100, %98 ve %96 olarak bulundu. Geliştirilen MAT ve ELISA ile 72 insan serumunda seropozitiflik oranı her iki teste de %4.2 olarak bulunurken, 190 koyun serumunda MAT ve ELISA seropozitifliği sırası ile %3.2 ve %4.7 olarak bulundu. Alınan sonuçlara göre ülke mizde tularemi insan ve hayvanlarda varlığını sürdüren bir infeksiyondur. Ancak daha sağlıklı epidemiyolojik yorum yapabilmek için daha çok sayıda seruma ve farklı hayvan türleri ile çalışılmasına ihtiyaç bulunmaktadır. Sonuç olarak, ELISA ve konfirme edici Western blotting kombinasyonunun tulareminin serolojik tanısında kullanılabilecek uygun bir kombinasyon olduğu düşünülmektedir.In this study, a microagglutination test (MAT) antigen stained by crystal violet and an in house Enzyme- Linked Immunosorbent Assay (ELISA) prototip were developed in order to detect antibodies against Francisella tularensis. Besides, Western blot technnique was used as the confirmatory test. According to the results, diagnostic sensitivity of MAT, ELISA and WB was 100%, while diagnostic specificities of these tests were 100%, 98% and 96%, respectively. Seropositivity rates of serum samples taken from 72 human were 4.2% for both tests. Seroposi tivity rates of 190 serum samples from sheep were 3.2% for MAT and 4.7% for ELISA. Results seem that tularemia exists in both humans and in animals. However, in order to make more definitive evaluation about the disease, more serum samples taken from various animal species are needed to be tested for this disease. As conclusion, it was thought that ELISA and confirmatory Western blot will be suitable combination for serologic diagnosis of tularemia

The first report of Brucella suis biovar 1 isolation in human in Turkey

SummaryBackgroundBrucella melitensis and B. abortus are the species generally isolated from human samples in Turkey. Several studies have also demonstrated the presence of antibodies against B. canis.Case report and studyBrucella spp. was isolated from blood culture from a 35-year-old male with clinical signs and symptoms of acute meningitis, including fever lasting for 1 week. Multiplex PCR demonstrated B. suis, and biochemical features indicated biovar 1.ConclusionsThis report is the first emphasizing that B. suis should be considered among the causes of brucellosis in Turkey

An Enzyme-Linked Immunosorbent Assay for Brucella Specific Antibody and Real-Time PCR for Detecting Brucella Spp. in Milk and Cheese in Sanliurfa, Turkey

The objective of this study was to investigate the presence of anti-Brucella antibody and Brucella spp. DNA in cow, sheep and goat milk and in Urfa cheese collected from markets and bazaars in Sanliurfa, located in southeast of Turkey. A total of 258 samples consisting of 178 raw milk (48 cow milk, 65 sheep milk and 65 goat milk) samples and 80 Urfa cheese samples were investigated. Anti-Brucella antibody was detected by indirect ELISA (i-ELISA), and the presence of Brucella spp. DNA was screened by real time Polymerase Chain Reaction (RTPCR). 16.6% of the cow, 6.1% of the goat and 6.1% of the sheep milk and 16.25% of the cheese samples were found as positive for brucella antibodies by i-ELISA. The RT-PCR assay amplified Brucella DNA from 18.75, 7.6 and 6.1% cow, goat and sheep milk samples respectively. Brucella DNA was amplified from 22.5% cheese samples. The 11.2% and 13.9% of the samples were found as positive by i-ELISA and RT-PCR respectively. This study indicates that milk and milk products consumed in Sanliurfa poses a risk to public health in terms of brucellosis. The combining usage of both i-ELISA and RT-PCR methods could lead to more reliable results to detect anti-Brucella antibody and Brucella spp. DNA from milk and cheese samples. (C) 2016 PVJ. All rights reserve

Vorhandensein von Staphylococcus aureus, Staphylokokken Enterotoxinen und ­antimikrobielle Resistenzen in traditionell hergestelltem Rohmilchkäse

The objectives of this study was to investigate the presence of Staphylococcus aureus, distribution of classical staphylococcal enterotoxin (SE) SEA to SEE, relevant gene/s and antimicrobial resistance pattern of S. aureus isolated from traditionally produced raw milk cheeses. A total of 106 fresh white cheese samples were examined. The 25 (23.6 %) of 106 cheese samples were found to be contaminated with coagulase positive staphylococci (CPS). From 52 isolates identified as S. aureus, one or more SEs was detected in 38.4 % of the isolates by ELISA whereas one or more se genes were detected in 50 % of the isolates by RT PCR. SEE (75 %) and see gene (61.5 %) were detected most frequently, whereas SED and sed gene were not detected in any isolates. Overall, 63.5 % of isolates were resistant to antimicrobial agents with 59.6 %, 13.5 %, 5.8 %, 5.8 % and 3.8 % of the isolates were resistant to penicillin, erythromycin, tetracycline, cefoxitin and kanamycin, respectively. The results of this study have revealed that cheeses made from raw milk were highly contaminated with S. aureus, therefore, creates a risk for public health due to the presence of enterotoxins as well as resistant strains against antimicrobial agents