Investigation of vector-borne diseases in dogs

In this study, a total of 186 blood samples were collected from kennel dogs consisting of 104 male and 82 female in five provinces (Mersin, Adana, Hatay, Gaziantep and Batman) of Turkey, and evaluated using molecular methods for the presence of canine vector-borne diseases (CVBDs). Overall, 10.8% of the sampled dogs were found to be infected with one or more CVBD pathogens investigated. Ehrlichia canis (17/186; 9.1%) was the most common CVBD pathogen, followed by Babesia canis vogeli (5/186; 2.7%) and Hepatozoon canis (1/186; 0.5%), respectively. Co-infection of E. canis with B. caniswas detected in 3 (1.6%) dogs. Infection with Rickettsia spp., Coxiella burnetii, Borrelia burgdorferi s.l., Francisella tularensis, Bartonella spp., Leishmania spp., Diroflaria immitis, Diroflaria repens, and Acanthocheilonema reconditum were not detected. No sex association with CVBDs was determined (p>0.05). The result of the study indicates the presence of three CVB pathogens, including the first report of B. canis and H. canis in the studied provinces.Bu çalışmada, Türkiye'nin beş farklı ilindeki (Mersin, Adana, Hatay, Gaziantep ve Batman) köpek barınaklarındanalınan 186 (104'ü erkek ve 82'si dişi) kan örneği vektör kaynaklı nakledilen patojenler yönünden moleküler yöntemlerlearaştırıldı. İncelenen örneklerin %10.8'inin en az bir veya birden fazla patojen ile enfekte olduğu tespit edildi. Ehrlichiacanis (17/186; %9.1) en yaygın vektör aracılı nakledilen patojen olup, bunu sırasıyla Babesia canis vogeli (5/186; %2.7) veHepatozoon canis (1/186; %0.5) izledi. E. canis ve B. canis ortak enfeksiyonu 3 (%1.6) köpekte tespit edildi. Rickettsia spp.,Coxiella burnetii, Borrelia burgdorferi s.l., Francisella tularensis, Bartonella spp., Leishmania spp., Diroflaria immitis,Diroflaria repens ve Acanthocheilonema reconditum enfeksiyonu saptanmadı. Vektör aracılı nakledilen patojenler yönündenpozitif bulunan köpeklerde yaş ve cinsiyet yönünden istatistiksel olarak önemli bir fark belirlenmedi (p> 0.05). Çalışılanillerde köpeklerde vektör aracılı nakledilen patojenlerden üçünün varlığı gösterilmiş ve çalışılan illerde ilk kez B. canis ve H.canis varlığı tespit edilmiştir

MAY BLASTOCYSTIS HOMINIS BE A POTENTIAL HAZARD FOR AN INTESTINAL DISEASE?

Blastocystis hominis (B. hominis)'in sürüngenlerde, kuşlarda, çeşitli memeli türlerinde ve böceklerde varlığı tanımlanmaktadır. Bu protozoon insanların bağırsaklarında bulunur. B. hominis'in vakuoler, granuler, ameboid, kist olmak üzere dört formu bulunmaktadır. Blastocystosis prevalansı, gelişmekte olan ülkelerde %30-%50, gelişmiş ülkelerde %1,5- %10'dur. Genellikle seyahat öyküsü olanlarda daha sık görülür. B. hominis enfeksiyonunun esas belirtileri, ishal ve karın ağrısı olmakla beraber, bulantı, iştahsızlık, kusma, kilo kaybı, halsizlik, baş dönmesi, gaz sancısı gibi tipik olmayan gastrointestinal şikayetler de görülür. Bu enfeksiyonun belirtileri kesin olmamasına rağmen, B. hominis‘in bağırsaklarda yangıya neden olarak patojenik rol oynadığı fikri ileri sürülmektedir. Günümüzde blastocystosis tedavisinde kemoterapötik ajan olarak metronidazol tercih edilmektedir. Blastocystis hominis (B. hominis) has been described in a variety of mammals, birds, reptiles, and even insects. The protozoon is commonly found in the intestinal tract of humans. Morphology of B. hominis has four major forms; vacuolar, granular, ameboid and cyst. The prevalence of Blastocystosis in humans appears to be higher in developing countries (30% to 50%) than in developed countries (1.5% to 10%), and has been associated with travel. Symptoms of B. hominis infection are mainly diarrhea and abdominal pain as well as nonspecific gastrointestinal symptoms such as nausea, anorexia, vomiting, weight loss, lassitude, dizziness, and flatulence. The significance of this human infection is uncertain although one idea has suggested a pathogenic role of B. hominis in causing intestinal inflammation. At present, the first choice of chemotherapeutic agent is metronidazole as described

Evaluation of symptomatic patients with resistant discharge

The aim of this study was to detect the presence of Chlamydia trachomatis, Neisseria (N.) gonorrhoeae, Mycoplasma (M.) hominis, M. genitalium, Ureaplasma (U.) urealyticum, and Trichomonas (T.) vaginalis in patients with resistant discharge. The study also evaluated the concordance of the diagnostic tests. Samples from 156 patients were tested by direct microscopy and culture for T. vaginalis and Mycoplasma IES for M. hominis and U. urealyticum. Multiplex Polymerase Chain Reaction (PCR) was used to determine the presence of six agents. Statistical analyses were performed using the SPSS program. Out of 156 patients, 38 had positive result for the agents tested. Of these 38 patients, 28 (73.7%) had single agent positivity and 10 (26.3%) had multiple agent positivity. The detection rate of U. urealyticum, M. hominis, N. gonorrhoeae, C. trachomatis, T. vaginalis, M. genitalium specifically was 10.3%, 9.6%, 6.4%, 3.2%, 2.6%, 0.6% respectively. N. gonorrhoeae and U. urealyticum were the most common in male patients, while M. hominis and U. urealyticum were mostly found in female patients. Different methods used for detecting T. vaginalis were compared to find that interrater reliability was perfect for culture-direct microscopy (κ:0.85; P&lt;0.001) and also for culture-PCR (κ:0.89; P&lt;0.001). The interrater reliability was moderate (κ:0.53; P&lt;0.001) for PCR-Mycoplasma IES test for M. hominis and fair (κ:0.21; P&lt;0.007) for U. urealyticum. U. urealyticum and M. hominis were among the most commonly found sexually transmitted infections (STI) agents in patients with resistant discharge. Multiple agent positivity was high and should be kept in mind in every STI case. </p

Evaluation of symptomatic patients with resistant discharge

The aim of this study was to detect the presence of Chlamydia trachomatis, Neisseria (N.) gonorrhoeae, Mycoplasma (M.) hominis, M. genitalium, Ureaplasma (U.) urealyticum, and Trichomonas (T.) vaginalis in patients with resistant discharge. The study also evaluated the concordance of the diagnostic tests. Samples from 156 patients were tested by direct microscopy and culture for T. vaginalis and Mycoplasma IES for M. hominis and U. urealyticum. Multiplex Polymerase Chain Reaction (PCR) was used to determine the presence of six agents. Statistical analyses were performed using the SPSS program. Out of 156 patients, 38 had positive result for the agents tested. Of these 38 patients, 28 (73.7%) had single agent positivity and 10 (26.3%) had multiple agent positivity. The detection rate of U. urealyticum, M. hominis, N. gonorrhoeae, C. trachomatis, T. vaginalis, M. genitalium specifically was 10.3%, 9.6%, 6.4%, 3.2%, 2.6%, 0.6% respectively. N. gonorrhoeae and U. urealyticum were the most common in male patients, while M. hominis and U. urealyticum were mostly found in female patients. Different methods used for detecting T. vaginalis were compared to find that interrater reliability was perfect for culture-direct microscopy (κ:0.85; P&lt;0.001) and also for culture-PCR (κ:0.89; P&lt;0.001). The interrater reliability was moderate (κ:0.53; P&lt;0.001) for PCR-Mycoplasma IES test for M. hominis and fair (κ:0.21; P&lt;0.007) for U. urealyticum. U. urealyticum and M. hominis were among the most commonly found sexually transmitted infections (STI) agents in patients with resistant discharge. Multiple agent positivity was high and should be kept in mind in every STI case. </p

Detection of Microsporidium spp. and Cryptosporidium spp. in diarrheic stool samples, ıdentification of species by PCR

Amaç: Ülkemizde boyama yöntemi ile tespiti henüz standardize edilememiş olan Microsporidium spp.'nin Weber'in trichrome boyama yöntemi ile saptanması, ishalli dışkılarda Microsporidium spp. ve Cryptosporidium spp.'nin tanısında mikroskobi ve PCR yöntemlerinin karşılaştırılması ve her iki parazitin tür ayrımının yapılmasını hedefledik. Gereç-Yöntem: Ekim 2005- Ocak 2009 tarihleri arasında, Dokuz Eylül Üniversitesi Tıp Fakültesi'nin çeşitli kliniklerine başvuran ve İzmir'in Buca ilçesine ait bir köyde saptanan ishal salgınından elde edilen 162 ishalli dışkı örneği çalışma kapsamına alındı. Tüm örnekler Cryptosporidium spp.'nin saptanması için Kinyoun acid fast ve Microsporidium spp.'nin saptanması için Weber'in trichrome boya yöntemleri ile mikroskobik olarak değerlendirildi. Ayrıca PCR yönteminin geçerliliğinin belirlenebilmesi için her iki parazite bu yöntem uygulandı. Bir sonraki aşamada Cryptosporidium ve Microspordium spp'nin tür ayrımlarının yapılabilmesi için RFLP yöntemi uygulandı. Bulgular: Kinyoun acid fast boya yöntemi ile 18 olgunun dışkı örneğinde Cryptosporidium spp. ookistleri saptandı. PCR yöntemi ile bu 18 olgudan 15'inde ve boya yöntemi ile negatif olarak değerlendirilen 6 olguda Cryptosporidium spp.'ye özgü bant saptandı. PCR ile parazit saptanan olgulara RFLP yöntemi uygulandı. Olgulardan birinde Cryptosporidium meleagridis'e, 20 olgunun dışkı örneğinde ise Cryptosporidium parvum'a özgü bantlar saptandı. Weber'in trichrome boya yöntemi ile 4 olgunun dışkı örneğinde Microsporidium spp. sporları saptandı. PCR yöntemi ile bu olgulardan sadece birinde Microsporidium spp.'ye özgü bantlar saptandı. PCR amplifikasyonunda tüm türlere yönelik primerler olan "panmicrosporidian primerler" kullanılarak tür ayrımı yapıldı. PCR ile parazit saptanan olgunun dışkı örneğinde Encephalitozoon intestinalis'e özgü bantlar saptandı. Sonuç: Cryptosporidium spp.'nin tanısında PCR yönteminin önemli bir yeri olduğuna, özellikle parazitin yoğun olduğu dışkılarda paraziti saptama gücünün yüksek olduğuna karar verildi. Microsporidium spp.'nin saptanmasında ise boya yönteminin tanıda geçerliliğini koruduğu, uygun DNA ekstraksiyon prosedürünün oluşturulmasına yönelik moleküler çalışmaların yapılması gerektiği düşünüldü. Anahtar kelimeler: Cryptosporidium spp., Microsporidium spp., Kinyoun acid fast, Weber'in trichrome boyası, PCR-RFLP. Aim: Our objective was to use Weber's trichrome staining method in detection of Microsporidium spp. of which convenient staining method for diagnosis yet to be standardized in our country, and to compare effectiveness of microscopy and PCR methods in detection of Microsporidium spp. and Cryptosporidium spp. in diarrheic stool samples and species identification of both parasites. Materials and Methods: Stool samples of 162 diarrheic patients who attended to various clinics of Hospital of Faculty of Medicine, University of Dokuz Eylul between October 2005 and January 2009 and of patients in an outbreak of diarrhea which took place in a village in Buca district of İzmir, were included in our study. All samples were stained by Kinyoun acid-fast method to search Cryptosporidium spp. and by Weber's trichrome staining method to search Microsporidium spp. and were examined by microscopy. Additionally, PCR was used for both parasites to test its validity as a diagnostic tool. In succeeding step, RFLP was used in species identification of both, Cryptosporidium spp. and Microspordium spp. Results: Oocysts of Cryptosporidium spp. were detected in 18 cases by Kinyoun acid fast staining method. PCR revealed Cryptosporidium spp. spesific bands in 15 of 18 patients who were staining positive and 6 of staining negative patients. RFLP method was applied to PCR positive stool samples and one sample had Cryptosporidium meleagridis spesific band while remaining 20 samples had Cryptosporidium parvum spesific bands. We observed spores of Microsporidium spp. in stool samples of 4 cases stained by Weber's trichrome method and only one of them was positive in PCR. Panmicrosporidian primers were used in amplification and PCR positive patient revealed Encephalitozoon intestinalis spesific bands. Conclusion: PCR is an important diagnotic tool in diagnosis of Cryptosporidium spp., especially in cases with high parasite density. In diagnosing Microsporidium spp., staining procedure sustains its validity and more molecular studies are needed to supply convenient DNA extraction procedure. Keywords: Cryptosporidium spp., Microsporidium spp., Kinyoun acid fast, Weber's trichrome, PCR-RFLP

MICROSPORIDIUM SPP. INFECTION IN AN IMMUNOCOMPROMISED CHILD DIAGNOSED BY POLYMERASE CHAIN REACTION

Microsporidium spp. may lead to a variety of clinical pictures like sinusitis, keratoconjunctivitis, hepatitis, myositis, peritonitis, nephritis, encephalitis and pneumonia in case of immune deficiencies. In this report, a case of diarrhea due to Microsporidium spp. has been presented. A four years old male patient who was followed with the diagnosis of myotonic dystrophia, was admitted to the hospital with the complaints of respiratory distress and fever. Due to the history of recurrent infections, further investigations was carried out to clarify the immunological status of the patient, and the total IgA and IgM levels were found as 14 mg/dl and 30 mg/dl, respectively (normal values were; 18-160 and 45-200 mg/dl, respectively). Following bronchoscopy done to enlighten respiratory distress, the patient developed high fever and watery diarrhea. Since bacteriological cultures of the stool yielded Shigella spp., antimicrobial therapy with ciprofloxacin was initiated. Parasitological examination of the stool done by Weber's modified trichrome dye, yielded Microsporidium spp. microscopically and albendazole was added to the treatment. Presence of Microsporidium spp. was confirmed by polymerase chain reaction with the use of Cl and C2 primers (Metabion, Germany) targeted to Microsporidium spp. and besides a 270 bp band specific for Encephalitozoon intestinalis was also obtained. This case emphasized that in case of diarrhea the stool samples of the immunocompromised patients should be evaluated in terms of Microsporidium spp. in addition to the routine parasitologic examinations

Detection and genotyping of cryptosporidium spp. in diarrheic stools by PCR/RFLP analyses

Aim: To compare microscopy and polymerase chain reaction (PCR) as methods for diagnosing Cryptosporidium spp. and identifying the genotype of the parasite in diarrheic stool

A WATERBORN PARASITE: CRYPTOSPORIDIUM

İlk kez 1907'de farelerde bildirilmiş olmasına rağmen, son 30 yıldır insanlarda hastalık yaptığı bilinmektedir. Moleküler biyolojik tekniklerdeki gelişmeler cryptosporidiosise bakış açımızı çok değiştirmiştir. Sadece immun yetmezlikli kişileri etkileyen nadir bir hastalık olarak görülürken, bugün su kaynaklı büyük salgınlardan sorumlu bir halk sağlığı sorunu olduğu anlaşılmıştır. Bu derlemede Cryptosporidium'a ait bilgiler son literatürler ışığında gözden geçirilmiştir. Although first described in mice in 1907, Cryptosporidium is known as a human disease for only 3 decades. Advances in moleculer biologic techniques has greatly changed our understanding of cryptosporidiosis. Previously known as a rare disease of immunodeficient patients, has became a public health concern responsible for waterborn outbreaks. In this review, we overviewed our knowledge of Cryptosporiodium in spite of current literature

Current Status in Intestinal Parasitic Infections: A Reference Laboratory Results

Objective: The aim of this study was to evaluate the results of examination for intestinal parasites in fecal samples sent to our laboratory by obtaining from patients applied to the hospital because of various complaints