Browsing by Author "Karakavuk, M"
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Item The Importance of the Contribution of Rapid Test, Serological and Molecular Methods in the Diagnosis of Two Imported Malaria Cases with Atypical MicroscopyZorbozan, O; Pullukçu, H; Sahar, EA; Karakavuk, M; Can, H; Tunali, V; Döskaya, M; Turgay, N; Töz, S; Özbilgin, AMalaria is a widespread and life-threatening disease in tropical and subtropical regions. In patients with typical clinical symptoms, malaria is considered as a preliminary diagnosis if there is a travel history to malaria-endemic areas. The basis of the laboratory diagnosis of malaria is the microscopic examination of Giemsa stained smears. On the other hand, the diagnosis and differentiation of Plasmodium species with microscopic examination may have some difficulties. In the first case, adifferent appearance from the classical Plasmodium vivax erythrocytic forms in infected erythrocytes were detected in 1% of all erythrocytes in thin smear blood preparations of a 26-year-old male with complaints of fever and chills and a story of travel to Nigeria. It was observed that parasitic nuclei were not prominent, and were located in the cytoplasm irregularly as chromatin or dye particles, nucleus fragments similar to Schuffner's granules in the form of scattered and granular spots were present in some erythrocytes, the cytoplasm of some Plasmodium erythrocytic forms were irregular and nuclei were not seen. There were no Schuffner's granules in any of the infected erythrocytes. P. vivax was detected by the rapid diagnostic test (OptiMAL, DiaMed GmbH, Switzerland), which searches for the antigens of Plasmodium species, in the peripheral blood sample of the patients. The P. vivax 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibodies against Plasmodium species were searched by using the Pan Malaria Antibody CELISA (CeLLabs Pty Ltd, Brookvale, Australia) kit in the patient's serum sample and the optical density (OD) value of the patient sample was measured five times the OD value of the positive control. In the second case, adifferent appearance from the classical P. falciparum erythrocytic forms in infected erythrocytes were detected in 12% of all erythrocytes in thin smear blood preparations of a 31-year-old male who has been suffering from persistent fever, severe headache, pain in the eyes and was known to be working in Nigeria. It was observed that some Plasmodium trophozoites have 1/3 of the size of erythrocytes such as P. vivax and have non-granular cytoplasm, some erythrocytic forms were round and the nucleus and cytoplasm were hardly distinguished, some of them were seen as crescent and close to the nucleus of the cytoplasm and some erythrocytic forms had characteristically a single nucleus and a scattered cytoplasm, similar to mature trophozoites of P. vivax. Although the Plasmodium young trophozoites were similar to P. vivax in means of magnitude, the forms in which the nuclei adhered to the erythrocyte wall were common. There were no P. falciparum gametocyte forms. P. falciparum like young trophozoite was observedonly in one of the four smears. P. falciparum was detected by the commercial rapid diagnostic test and P. falciparum 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibody formation against Plasmodium species was not detected in the ELISA test. In these case reports, the importance of the support of rapid diagnostic tests, serological and molecular methods to microscopic diagnosis and species determination of two imported malaria cases were demonstrated.Item Development of a Rapid-Crypto Colorimetric LAMP Test to Detect Cryptosporidiosis in Feces of Newborns CalvesKarakavuk, M; Can, H; Can, S; Karakavuk, T; Döskaya, M; Döskaya, ADBackgroundCryptosporidiosis is a disease that causes major intestinal damage in humans and animals. The causative agents of the disease are Cryptosporidium species. In newborn calves, diarrhea can lead to death, resulting in significant economic losses for the farms. Therefore, accurate, rapid, and cost-effective diagnosis of the disease is very important.Material and methodsIn this study, a novel colorimetric loop-mediated isothermal amplification (LAMP) test named Rapid-Crypto Colorimetric LAMP test targeting Cryptosporidium spp. 18S rRNA gene was developed to detect cryptosporidiosis in the feces of newborn calves. The analytical sensitivity of the test was determined by plasmid controls. Clinical sensitivity was determined using the feces of 127 calves collected from farms in Izmir and Manisa provinces. All of the samples were also investigated with Real-Time PCR targeting the Cryptosporidium spp. COWP gene. Cross-reactivity was tested using the DNA of other parasites and bacteria.ResultsAccording to the results, the analytical sensitivity of the Rapid-Crypto Colorimetric LAMP test was found as 1 copy plasmid/reaction. When the results were compared with the Real-Time PCR test, the sensitivity of the Rapid-Crypto Colorimetric LAMP test was 100% and the specificity was 97.4%. The test did not cross-react with other parasites and bacteria. ConclusionThe Rapid-Crypto Colorimetric LAMP test developed in this study provides an advantage in the diagnosis of Cryptosporidium spp. in calf stool samples since it can be applied in basic laboratories or in the field, does not require experienced personnel, and has high sensitivity. Moreover, diagnosis can be made with the naked eye without using any device.Item Molecular prevalence and genotypes of Enterocytozoon bieneusi in cancer patients under chemotherapy in Aegean region of TurkiyeGökmen, AA; Oner, TO; Alak, SE; Koçkaya, ES; Güvendi, M; Karabey, M; Alacacioglu, A; Pektas, B; Döskaya, AD; Karakavuk, M; Döskaya, M; Un, C; Gürüz, AY; Kaya, S; Can, HBackground Enterocytozoon bieneusi is the most common species found in humans. Although E. bieneusi has been investigated in humans, genotype profile of E. bieneusi is not known in Turkiye. Methods In this study, we screened E. bieneusi in patients (n = 94) with different types of malignant solid tumors by Real Time PCR and then sequenced E. bieneusi positive samples. All cancer patients were undergoing chemotherapy and had diarrhea. Moreover, as control groups, we also screened E. bieneusi in patients with diarrhea (n = 50) and without diarrhea (n = 50). Results Among all patients analyzed, 33 (17%) were found to be E. bieneusi-positive. As the patients were categorized, the molecular prevalence of E. bieneusi increased to 25.5% among cancer patients with diarrhea. However, the molecular prevalence of E. bieneusi was found to be lower in patients with presenting only diarrhea (8%) and patients without diarrhea (10%). The high molecular prevalence value detected among cancer patients with diarrhea was also statistically significant compared to other patient groups (P = 0.00112 and P = 0.0269). Among the 33 Real Time PCR positive samples, 10 of them were amplified by nested PCR and among these 10 samples, 6 of them were successfully genotyped. The phylogenetic tree showed the presence of D and Type IV which were also identified in stray cats living in & Idot;zmir in our previous study. Conclusions High molecular prevalence value indicates the importance of screening stool samples of cancer patients with diarrhea for E. bieneusi and genotyping results indicate that D and Type IV are circulating between humans and cats.Item Cryptosporidium spp. during chemotherapy: a cross-sectional study of 94 patients with malignant solid tumorKarabey, M; Can, H; Öner, TÖ; Döskaya, M; Alak, SE; Döskaya, AD; Karakavuk, M; Köseoglu, AE; Ün, C; Gürüz, AY; Alacacioglu, A; Pektas, B; Gül, A; Kaya, S; Gokmen, AABACKGROUND: Cryptosporidium spp. is a protozoan parasite that infects many vertebrate animals, including humans. Since Cryptosporidium spp. can cause chronic life-threatening diarrhea and severe malabsorption in immunocompromised patients, we investigated the prevalence of this parasite among patients undergoing chemotherapy for malignant solid tumors. OBJECTIVE: Investigate the prevalence of Cryptosporidium spp. in stool samples. DESIGN: Cross-sectional. SETTING: Tertiary care. PATIENTS AND METHODS: Stool samples were collected from adult patients with malignant solid tumors receiving chemotherapy and diarrhea. Cryptosporidium spp. prevalence was determined using Ziehl-Neelsen staining, ELISA, and real-time PCR targeting of the COWP gene. MAIN OUTCOME MEASURE: The prevalence of Cryptosporidium spp. in patients undergoing chemotherapy for malignant solid tumors. SAMPLE SIZE: 94 RESULTS: The prevalence was 2.1% (2/94), 5.3% (5/94), and 5.3% (5/94) as detected by Ziehl-Neelsen staining, real-time PCR and ELISA, respectively. The prevalence reached 8.5% (8/94) using all results obtained from the three methods. Among eight positive stool samples, four were positive by at least two different methods (Ziehl-Neelsen staining-ELISA or ELISA-real-time PCR) whereas the remaining four were positive by either ELISA or real-time PCR. CONCLUSION: These findings show the risk of cryptosporidiosis in cancer patients and the necessity to use at least two diagnostic methods during the diagnosis of cryptosporidiosis to reach more accurate and trustworthy results. LIMITATIONS: Further studies with a larger sample size are recommended. CONFLICT OF INTEREST: None.Item Investigation of the role of stray cats for transmission of toxoplasmosis to humans and animals living in Izmir, TurkeyKarakavuk, M; Can, H; Selim, N; Yesilsiraz, B; Atli, E; Sahar, EA; Demir, F; Gül, A; Özdemir, HG; Alan, N; Yalçin, M; Özkurt, O; Aras, M; Çelik, T; Can, S; Döskaya, AD; Gürüz, AY; Döskaya, MIntroduction: Toxoplasma gondii is a protozoan parasite that has a widespread distribution among mammalians and birds. One of the reasons for the high prevalence may be due to ingesting oocyst disseminated by stray cats' feces. In Turkey, most of the citizens are closely associated with stray cats and they love to pet and feed them on the streets. In this study, we aimed to determine the prevalence of T. gondii DNA in feces of stray cats living in Izmir, Turkey in order to identify the transmission potential to humans and other animals. Methodology: Feces and blood samples of 465 stray cats were investigated for the presence of T. gondii oocysts by microscopy and for the presence of T. gondii DNA by two real time PCR methods. Furthermore, serum samples were analyzed for anti-T. gondii IgG antibodies using an ELISA. Results: Oocysts were detected in 0.43% of the stray cats by microscopy. T. gondii DNA was detected in 14.37% of the stray cats' feces samples. The seroprevalence rate was 37.84%. In the feces and/or blood PCR positive group, 35.89% of them were seropositive. Among the 176 seropositive cats, T. gondii DNA was detected in feces of 27 cats (15.34%). Conclusions: This study first time showed the inter relation of T. gondii DNA in feces and blood samples and seropositivity. In sum, over 14% of the stray cats living outdoor may have an important role in transmission of toxoplasmosis to humans in Izmir as well as to other animals.Item Molecular prevalence of Blastocystis sp. and subtype diversity in fecal samples collected from cattle in dairy farms in TurkeyOner, TO; Karakavuk, M; Doskaya, AD; Guvendi, M; Gül, A; Koseoglu, AE; Alak, SE; Guruz, AY; Un, C; Doskaya, M; Can, HClose contact with infected animals is one of the main risk factors for zoonotic transmission of enteric protozoan parasite Blastocystis and thus, several animal species are being screened for the detection of the zoonotic subtypes. For this purpose, 22 fecal samples were collected from healthy cattle aged > 2 months and 39 fecal samples were also collected from cattle (aged <2 months) which are treated with TMP-SMX due to diarrhea. Later, Blastocystis sp. and subtypes were investigated by a PCR targeting the SSU rRNA gene and subsequently by sequencing. Among the 22 fecal samples collected from healthy cattle, Blastocystis was detected in 12 of them with a prevalence rate of 54.5 %. Among Blastocystis-positive samples, five different subtypes (ST3, ST5, ST10, ST12, and ST13) were detected. The predominant subtype was ST10 (allele 152) with a prevalence rate of 50 % (6/12). In the other group treated with TMP-SMX due to diarrhea, Blastocystis was detected in only one (2.56 %;1/39) fecal sample and its subtype was ST1 (allele 2). High prevalence of Blastocystis as well as predominance of ST10 (allele 152) were detected in healthy cattle. The identification of zoonotic ST1, ST3, ST5 and ST12 subtypes among the detected subtypes with a high prevalence (46.1 %; 6/13) showed the importance of cattle as a source for transmission of Blastocystis to humans. It was observed that the efficiency of TMP-SMX on the clearance of Blastocystis in cattle was very strong. Moreover, to our knowledge, this is the first study detecting Blastocystis ST13 subtype in the cattle.Item Prevalence of toxoplasmosis and genetic characterization of Toxoplasma gondii strains isolated in wild birds of prey and their relation with previously isolated strains from TurkeyKarakavuk, M; Aldemir, D; Mercier, A; Sahar, EA; Can, H; Murat, JB; Döndüren, O; Can, S; Özdemir, HG; Döskaya, AD; Pektas, B; Dardé, ML; Gürüz, AY; Döskaya, MToxoplasma gondii is a protozoon parasite that causes congenital toxoplasmosis, as well as other serious clinical presentations, in immune compromised humans. Analyses of the prevalence and genotyping of strains from the definitive host and intermediate hosts will help to understanding the circulation of the different strains and elucidating the role of the genotype (s) in human toxoplasmosis. Turkey has a specific geographic location bridging Africa, Europe, and Asia. We hypothesized that T. gondii strains may have been transferred to Turkey from these continents via migratory birds or vice versa. The present study aimed to assess the prevalence of toxoplasmosis in wild birds of prey of Izmir and Manisa provinces as well as genetically characterize T. gondii strains from these wild birds to show the relation between bird strains and neighboring stray cats as well as human strains previously isolated in Turkey. Tissues obtained from 48 wild birds were investigated for the presence of T. gondii DNA and then bioassayed in mouse. Isolated strains were genotyped using 15 microsatellite markers. The prevalence of T. gondii DNA was found to be 89.6% (n: 43/48) in wild birds. Out of 43 positive samples, a total of 14 strains were genotyped by 15 microsatellite markers. Among them, eight were type II, three were type III and three were mixture of genotypes (two type II/II and one was II/III). These are the first data that showed the presence of T. gondii and types II and III genotypes in wild birds of Turkey. Moreover, Africa 1 was not detected. In addition, cluster analysis showed that T. gondiistrains within type II and III lineage have close relation with strains previously isolated from stray cats in Izmir. Further studies are required to isolate more strains from human cases, other intermediate hosts, and water sources to reveal this relation.Item Molecular investigation of Blastocystis sp. and its subtypes in cancer patients under chemotherapy in Aegean region, TurkeyOner, TO; Karabey, M; Can, H; Doskaya, AD; Karakavuk, M; Gul, A; Koseoglu, AE; Doskaya, M; Un, C; Guruz, AY; Kaya, S; Pektas, B; Gokmen, AABlastocystis sp. is a common enteric protist found in humans and many other animals. Although the clinical relevance of Blastocystis sp. is currently fully unknown for humans, the prevalence of Blastocystis and subtypes are investigated in immunocompetent individuals presenting with symptoms like diarrhea or immunocompromised individuals including cancer patients. In this comprehensive study, the prevalence of Blastocystis sp. and subtypes were investigated in patients (n=94) with different types of malignant solid tumors using PCR targeting SSU rDNA gene and sequencing. All patients were undergoing chemotherapy and had diarrhea. According to obtained results, 46 patients were found to be Blastocystis positive and the molecular prevalence was detected as 48.9%. Among the positive specimens, 43 (43/46; 93.5%) of them were successfully subtyped. ST4 was the most predominant subtype and detected in 24 (55.8%) patients, followed by ST1 (11 patients, 25.6%) and ST3 (8 patients, 18.6%). In the colon cancer group, which had the highest number of patients, Blastocystis sp. was detected with a higher prevalence rate of 61.5% compared with the prevalence rate (48.9%) of all patients. Interestingly, ST3 was not detected in any of this patient group in contrast to ST4 and ST1. In conclusion, high prevalence of the Blastocystis in the immunocompromised patient groups shows the susceptibility of this patient group against any other infectious agents.