The role of CD1a expression in the diagnosis of cutaneous leishmaniasis, its relationship with leishmania species and clinicopathological features.

Cutaneous leishmaniasis is caused by a flagellated protozoan transmitted by the bite of a female sandfly. The clinical and demographic details of this disease, predominantly affecting immunocompetent individuals, are recognized by the WHO as a Neglected Tropical Disease. We sought to determine the usability of CD1a immunohistochemical staining to detect amastigotes especially in cases where leishmaniasis is suspected but evident amastigotes could not observed. We also evaluated the relationship between CD1a expression and leishmania subtypes. A total of 84 cases diagnosed with leishmaniasis or suspected leishmania on histo-morphological evaluation of skin biopsies were included in the study. Amastigotes were easily detected in hematoxylin eosin in 18 of 84 cases. In 23 cases, amastigotes could not detect in hematoxylin eosin sections. The immunostains for CD1a are demonstrated amastigotes in 60 of 84 cases. However, a small number of amastigotes became visible by positive staining with CD1a in 43.4% of the cases in that amastigotes could not detected in hematoxylin eosin. A statistically significant correlation was found between amastigote amount in hematoxylin eosin and CD1a expression. In addition, a significant correlation was observed between CD1a expression, age and clinical pre-diagnosis of the cases. It was observed that amastigotes were easily detected in hematoxylin eosin in Leishmania Infantum / donovani positive cases in polymerase chain reaction (PCR), and at the same time, it was found that CD1a expression was significantly higher. Using histopathology examination with CD1a staining and/or PCR methods, a diagnosis of leishmaniasis can be established and early treatment initiated. This contributes to reduce transmission and prevalence.

[Investigation of the presence of Cryptosporidium spp. in different water sources in Mersin province, Turkey]. / Mersin Ilinde Farkli Su Kaynaklarinda Cryptosporidium spp. Varliginin Arastirilmasi

Cryptosporidium is an intracellular protozoon that causes enteritis in human and animals. Contaminated water and food are the major sources for the transmission of oocysts via oral-fecal route. It is reported that the prevalence of cryptosporidiosis is higher in developing countries than developed countries because of inefficient sanitation and disinfection facilities for drinking water. The most frequently detected species is Cryptosporidium parvum leading to high morbidity in healthy subjects and also fatal infections in immunocompromised patients. The acid-fast staining method is widely used in the diagnosis of cryptosporidiosis. Nowadays, Cryptosporidium could easily be detected in water supplies and asymptomatic carriers by molecular techniques to obtain epidemiological data. In this study it was aimed to detect and identify Cryptosporidium oocysts in different water sources in Mersin province, Turkey. A total of 135 water samples (70 taps, 50 wells and 15 sewage) collected from city center (n= 25) and from Tarsus (n= 32), Mezitli (n= 33) and Karaduvar (n= 45) counties between March 2007 and May 2009 were included in the study. Water samples in 10 liter volumes, were filtered by 0.45 µm pore-sized membrane filter vacuum/ pressure pumping technique. Cryptosporidium oocysts in filtrates were detected by modified cold Kinyoun acid-fast stain (MCK) technique and also identified and typed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. MCK yielded three and PCR yielded seven positive results. All the strains were identified as C.parvum by PCR-RFLP method. All of the three MCK-positive samples were also found positive with PCR, however four PCR positive samples were MCK-negative. Thus, the prevalence of C.parvum was estimated as 5.2% (7/135) in our region. Of seven positive samples, one was a sewage water sample collected from the city center, while the remaining (two tap water, two well water and two sewage water samples) belonged to the samples collected from Karaduvar county, interestingly. It was thought that deficient infrastructure and use of well water as drinking water supply in Karaduvar region might be the cause of high rate of Cryptosporidium (6/45; 13.3%). Further studies which will determine the genotypes and investigate the phylogenetic relationship between these Cryptosporidium spp., might aid to the epidemiology of cryptosporidiosis in our region.

[Isolation and identification of Mycobacterium tuberculosis from the urine samples by conventional and molecular methods]. / Konvansiyonel ve moleküler yöntemler ile idrar örneklerinden Mycobacterium tuberculosis izolasyonu ve tanimlanmasi.

Genitourinary tuberculosis presents a challenge in diagnosis and treatment due to variations in clinical and radiological signs, insufficient patient history and difficulty in the isolation of the bacilli. The aim of this study was to isolate and identify Mycobacterium tuberculosis from the urine samples obtained from patients with suspected urinary tuberculosis admitted to our hospital by using Ehrlich-Ziehl-Neelsen (EZN), culture and polymerase chain reaction-restriction analysis (PCR-RFLP) methods. A total of 1004 urine samples collected from 437 patients who were admitted to our hospital between January 2004-July 2006, were inoculated on Löwenstein-Jensen (LJ) and/or BACTEC 12B (Becton Dickinson, USA) after decontamination and, direct preparations stained with EZN method were evaluated microscopically. M. tuberculosis complex (MTC) and mycobacteria other than tuberculosis (MOTT) were differentiated by nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) test and the susceptibility testing for the MTC strains to primary antituberculosis drugs were performed by BACTEC 460 TB (Becton Dickinson, USA) system. PCR-RFLP method was performed for the identification of Mycobacterium spp. Twenty-two (5%) patients have yielded positive results by at least one of the conventional methods (EZN, LJ and/or BACTEC). Fifteen samples were positive for acido-resistant bacilli (ARB) by EZN method, and 17 samples were positive for mycobacterial growth in the cultures. Ten of 22 patients were found positive by both of the methods, while seven were culture positive but ARB negative and five were culture negative but ARB positive. These five patients received BCG treatment because of the presence of bladder tumor. Twelve (70.5%) of 17 strains isolated from culture were identified as MTC, while five (29.4%) were identified as M. fortuitum. Of 12 MTC isolates, eight (66.7%) were found susceptible to all of the antituberculosis agents, while one was found resistant to isoniazide (INH) and ethambutole (ETB), one was resistant to INH and rifampicin (RIF), and two were resistant to only INH. It is concluded that, in order to identify mycobacteria and to perform antituberculous susceptibility tests, cultivation of mycobacteria is a prerequisite.