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Öğe Cryptosporidium spp. during chemotherapy: a cross-sectional study of 94 patients with malignant solid tumor(K Faisal Spec Hosp Res Centre, 2021) Karabey, Mehmet; Can, Huseyin; Oner, Tulay Oncu; Doskaya, Mert; Alak, Sedef Erkunt; Doskaya, Aysu Degirmenci; Karakavuk, MuhammetBACKGROUND: 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.Öğe Development of a hexavalent recombinant protein vaccine adjuvanted with Montanide ISA 50V and determination of its protective efficacy against acute toxoplasmosis(Bmc, 2020) Sahar, Esra Atalay; Can, Huseyin; Iz, Sultan Gulce; Doskaya, Aysu Degirmenci; Kalantari-Dehaghi, Mina; Deveci, Remziye; Doskaya, MertBackgroundToxoplasma gondii is an obligate intracellular parasite that can infect almost all warm-blooded animals, avian species and humans. Toxoplasmosis is asymptomatic in healthy individuals, whereas it may lead to death in immune suppressed or deficient patients. A vaccine against T. gondii is required to prevent consequences of the infection. the aim of this study is to generate a multivalent recombinant protein vaccine against T. gondii.Methods49 previously discovered antigenic proteins of T gondii were evaluated by their expression level in E. coli and by comprehensive bioinformatics analyses to determine antigenic epitopes. Based on these analyses, six vaccine candidate proteins were selected to generate a hexavalent recombinant protein vaccine adjuvanted with Montanide ISA 50V. Humoral and cellular immune responses were determined by flow cytometry and ELISA. Vaccinated mice were challenged with T. gondii Ankara strain tachyzoites.ResultsIn mice vaccinated with hexavalent vaccine, strong total IgG (P<0.0001) and IgG2a (P<0.001) responses were induced compared to controls, the ratio of CD4(+) and CD8(+) T lymphocytes secreting IFN-gamma increased, and significantly higher extracellular IFN-gamma secretion was achieved compared to the controls (P<0.001). the survival time of the vaccinated mice increased to 8.382.13days which was significantly higher than controls (P<0.01).Conclusions Altogether, these results show that the hexavalent vaccine which is developed for the first time against T. gondii induced strong and balanced Th1 and Th2 immune responses as well as conferred significant protection against challenge with lethal toxoplasmosis in murine model.Öğe Development of a Rapid-Crypto Colorimetric LAMP Test to Detect Cryptosporidiosis in Feces of Newborns Calves(Springer Int Publ Ag, 2024) Karakavuk, Muhammet; Can, Huseyin; Can, Sengul; Karakavuk, Tugba; Doskaya, Mert; Doskaya, Aysu DegirmenciBackgroundCryptosporidiosis 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.Öğe Diagnostic Value of a Rec-ELISA Using Toxoplasma gondii Recombinant SporoSAG, BAG1, and GRA1 Proteins in Murine Models Infected Orally with Tissue Cysts and Oocysts(Public Library Science, 2014) Doskaya, Mert; Caner, Ayse; Can, Huseyin; Iz, Sultan Gulce; Gedik, Yaprak; Doskaya, Aysu Degirmenci; Kalantari-Dehaghi, Mina; Guruz, YukselToxoplasma gondii causes congenital toxoplasmosis in newborns resulting with fetal anomalies. Determining the initiation time of infection is very important for pregnant women and current serological assays have drawbacks in distinguishing the recently acute toxoplasmosis. Diagnosis of recently acute infection may be improved by using stage specific antigens in serological assays. In the present study, the diagnostic value of sporozoite specific SporoSAG, bradyzoite specific BAG1 proteins and GRA1 protein expressed by all forms of the parasite have been evaluated ELISA using sera systematically collected from mice administered orally with tissue cyst and oocysts. The anti-SporoSAG IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 1, 10, and 15 (P<0.01). The anti-BAG1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly at days 15, 40, and 120 (P<0.05). The anti-GRA1 IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 2, 10, and 40 (P<0.01). The anti-GRA1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly only at day 40 (P<0.05). The anti-SporoSAG, anti-BAG1, and anti-GRA1 IgG titers of mice showed significant increases at day 40 (P<0.05) and decrement started for only anti-GRA1 IgG at day 120. The presence of anti-SporoSAG IgM and IgG antibodies can be interpreted as recently acute infection between days 10-40 because IgM decreases at day 40. Similarly, presence of anti-BAG1 IgM and absence of IgG can be evaluated as a recently acute infection that occurred 40 days before because IgG peaks at day 40. A peak in anti-GRA1 antibody level at first testing and reduction in consecutive sample can be considered as an infection approximately around day 40 or prior. Overall, recombinant SporoSAG, BAG1 and GRA1 proteins can be accepted as valuable diagnostic markers of recently acute toxoplasmosis.Öğe Discovery of new Toxoplasma gondii antigenic proteins using a high throughput protein microarray approach screening sera of murine model infected orally with oocysts and tissue cysts(Bmc, 2018) Doskaya, Mert; Liang, Li; Jain, Aarti; Can, Huseyin; Iz, Sultan Gulce; Felgner, Philip Louis; Doskaya, Aysu Degirmenci; Davies, David Huw; Guruz, Adnan YukselBackground: Toxoplasma gondii is an obligate intracellular protozoan parasite that causes congenital toxoplasmosis, as well as other serious clinical presentations in immune compromised humans. The parasite has also been recently linked to behavioral diseases in humans and other mammalian hosts. New antigens are being evaluated to develop a diagnostic kit for the diagnosis of acute infection or a protective vaccine. Methods: In this study, we have focused on the discovery of new antigenic proteins from T. gondii genomic data using a high throughput protein microarray screening. To date, microarrays containing > 2870 candidate exon products of T. gondii have been probed with sera collected from patients with toxoplasmosis. Here, the protein microarrays are probed with well-characterized serum samples from animal models administered orally with oocysts or tissue cysts. The aim was to discover the antigens that overlap in the mouse profile with human antibody profiles published previously. For this, a reactive antigen list of 240 antigens recognized by murine IgG and IgM was identified using pooled sera from orally infected mice. Results: Analyses of screening data have identified plenty of antigens and showed strong immunogenicity in both mouse and human antibody profiles. Among them, ROP1, GRA2, GRA3, GRA4, GRA5, GRA6, GRA7, GRA8, GRA14, MIC1, MIC2 and MAG1 have shown strong immunogenicity and used as antigen in development of vaccines or serological diagnostic assays in previous studies. Conclusion: In addition to the above findings, ROP6, MIC12, SRS29A and SRS13 have shown strong immunogenicity but have not been tested in development of a diagnostic assay or a vaccine model yet.Öğe Discovery of new Toxoplasma gondii antigenic proteins using a high throughput protein microarray approach screening sera of murine model infected orally with oocysts and tissue cysts (vol 11, 393, 2018)(Bmc, 2024) Doskaya, Mert; Liang, Li; Jain, Aarti; Can, Huseyin; Iz, Sultan Gulce; Felgner, Philip Louis; Doskaya, Aysu Degirmenci[Abstarct Not Available]Öğe Genetic Characterization of Toxoplasma gondii Isolates and Toxoplasmosis Seroprevalence in Stray Cats of Izmir, Turkey(Public Library Science, 2014) Can, Huseyin; Doskaya, Mert; Ajzenberg, Daniel; Ozdemir, H. Gokhan; Caner, Ayse; Iz, Sultan Gulce; Doskaya, Aysu Degirmenci; Atalay, Esra; Cetinkaya, Cagdas; Urgen, Saygun; Karacali, Sabire; Un, Cemal; Darde, Marie-Laure; Guruz, YukselCurrently, some Toxoplasma gondii genotypes are being associated with serious clinical presentations. A recent report showing the Africa 1 genotype in two local congenital toxoplasmosis cases acquired in Turkey formed the basis of this study because atypical Africa 1 genotype is most frequently detected in animals and patients from sub-Saharan Africa. Since stray cats are considered as the linkage between wild life and urban life in T. gondii transmission, the present study aimed to isolate and characterize T. gondii strains circulating in stray cats of Izmir (Western Turkey). A secondary objective was to determine toxoplasmosis seroprevalence in this cat population. Tissues obtained from 100 deceased stray cats were bioassayed and isolated strains were genotyped using 15 microsatellite markers. In addition, toxoplasmosis seroprevalence was analyzed in 1121 cat sera collected from several large veterinary clinics in Izmir. Among the 22 isolates, 19 were Type II (86.3%), two were Type III (9%) and one was Africa 1 genotype (4.5%). The overall seropositivity rates in cats were 42-48% and 33.4-34.4% according to IFA and ELISA, respectively. Seroprevalence in deceased cats was significantly higher than in healthy cats (P = 0.0033). Finding both the major clonal Type II lineage together with the Type III lineage also found in Middle East, and an atypical genotype, Africa 1 appears consistent with the specific geographic location of Turkey between three continents and raises the possibility of transportation of these strains between continents through trade routes or long distance migratory birds. In addition, the first large study of toxoplasma seroprevalence in a stray cat population was also reported. The relatively high seropositivity rates and the variety of T. gondii genotypes confirm the local stray cat population as a risk factor for human toxoplasmosis in Izmir.Öğe Genotyping of Enterocytozoon bieneusi isolates detected in stray cats of Izmir, Turkiye(Springer, 2023) Sügeç, Ecem; Güvendi, Mervenur; Karakavuk, Muhammet; Alak, Sedef Erkunt; Doskaya, Aysu Degirmenci; Un, Cemal; Doskaya, MertThe phylum Microsporidia includes obligate intracellular parasites that can infect humans and various animals. To date, 17 different species within the phylum have been reported to infect humans. Among them, Enterocytozoon bieneusi (E. bieneusi) is one of the most frequently detected species in humans. Identification of E. bieneusi as well as its genotypes in humans and animals is important to reveal their role in transmission to each other. Cats are blamed as the source of E. bieneusi transmission to humans. In this study, we aimed to genotype 170 E. bieneusi positive samples isolated from stool of stray cats living in Izmir province of Turkiye. According to the results, 47 samples were amplified by nested PCR protocol targeting ITS region and successfully sequenced. The phylogenetic analysis showed the presence of zoonotic genotype D and type IV in stray cats, which are also frequently detected in humans. Among the E. bieneusi genotypes detected, the prevalence of type IV (93.6%; 44/47) was very high compared to genotype D. Overall, the identification of zoonotic genotypes of E. bieneusi supports that stray cats can play an important role in the transmission of E. bieneusi to humans in Izmir.Öğe Genotyping ofPneumocystis jiroveciiisolates obtained from clinical samples by multilocus sequencing: a molecular epidemiology study conducted in Turkey(Springer, 2020) Surgec, Ecem; Can, Huseyin; Doskaya, Mert; Karakavuk, Muhammet; Sahar, Esra Atalay; Doskaya, Aysu Degirmenci; Demir, SamiyePneumocystis jiroveciiis an opportunistic respiratory pathogen causing Pneumocystis pneumonia (PcP) in immunocompromised patients. the aim of this study was to investigate the genetic diversity ofP. jiroveciiisolates (n: 84) obtained from PcP patients using multilocus sequencing method based on mt26S, SOD, and CYB loci. Among the 84 clinical samples that were positive forP. jiroveciiDNA, 31 (36.90%) of them were genotyped using at least one locus. of the 31 clinical samples, 26 of them were successfully genotyped using all loci whereas three samples were genotyped using either mt26S/CYB loci or mt26S/SOD loci. Additionally, there were two more clinical samples that were genotyped using CYB or SOD locus. Using mt26S locus, genotypes 2, 3, 7, and 8 were detected. Frequencies of genotype 7 and 8 were higher and both of them were found in 11 (n: 29; 37.93%) clinical samples. Using SOD locus, SOD 1, 2, and 4 genotypes were detected. SOD 1 was the predominant genotype (20/28; 71.42%). During the analyses of CYB locus, CYB 1, 2, 5, 6, and 7 as well as a new CYB genotype were detected. CYB 1 (16/29; 55.17%) and 2 (10/29; 34.48%) were the predominant genotypes. Overall, according to the multilocus sequencing results E, F, M, N, P, and V multilocus genotypes were detected among the PcP patients. in addition, SOD 1 was the predominant genotype and CYB had a more polymorphic locus.Öğe GRA8 DNA vaccine formulations protect against chronic toxoplasmosis(Academic Press Ltd- Elsevier Science Ltd, 2021) Karakavuk, Muhammet; Can, Huseyin; Gul, Aytul; Doskaya, Aysu Degirmenci; Alak, Sedef Erkunt; Un, Cemal; Guruz, Adnan YukselToxoplasma gondii has a very wide host range and infects all warm-blooded animals including humans. The disease causes great economic losses both in animals and humans. Vaccination is the most effective approach to fight against toxoplasmosis however an effective vaccine has not been developed yet. In the present study, GRA8 protein of T. gondii that showed high immunogenicity in our previous microarray screening study was used to develop a DNA vaccine using pcDNA 3.3 vector for the first time. In order to increase the potency of the DNA vaccine, 10 times lower amount of GRA8 DNA vaccine was combined with molecular adjuvant CpG and formulated into a commercial liposome (pcDNA3.3-GRA8+CpG+Escort). Mice were vaccinated intramuscularly two times at three-week intervals and challenged orally with the T. gondii PRU strain tissue cysts. The humoral immune response was determined by Western Blot and ELISA. The cellular immune response was analyzed by flow cytometry, cytokine ELISA and MTT assay. Among the vaccine groups, pcDNA3.3-GRA8 and pcDNA3.3-GRA8+CpG+Escort induced strong IgG response compared to controls (P < 0.001). The IgG1 and IgG2a responses showed a balanced Th1-Th2 polarization. The ratio of CD4(+) and CD8(+) T lymphocytes secreting IFN-gamma increased, and significantly higher extracellular IFN-gamma secretion was achieved compared to the controls (P < 0.01). The amount of tissue cysts in the group of mice vaccinated with pcDNA3.3-GRA8 decreased significantly compared to control groups (P < 0.0001). In the group vaccinated with pcDNA3.3-GRA8+CpG+Escort, the amount of tissue cysts also decreased significantly compared to PBS (P = 0.0086) and Empty plasmid+CpG+Escort (P = 0.0007) groups. This study showed for the first time that pcDNA 3.3. vector encoding GRA8 with or without CpG and Liposome can induce strong cellular and humoral immune responses and confer strong protection against mouse model of chronic toxoplasmosis.Öğe The molecular and serological investigation of Feline immunodeficiency virus and Feline leukemia virus in stray cats of Western Turkey(Elsevier Sci Ltd, 2021) Muz, Dilek; Can, Huseyin; Karakavuk, Muhammet; Doskaya, Mert; Ozdemir, Huseyin Gokhan; Doskaya, Aysu Degirmenci; Sahar, Esra AtalayThis study aimed to investigate the Feline immunodeficiency virus (FIV) / Feline leukemia virus (FeLV) infection prevalence among looking healthy stray cats in Western Turkey by serologic and molecular-based tests. A total of 1008 blood samples from the stray cats were used in this study. All samples were tested for FIV antibodies / proviral DNA and FeLV antibodies / antigens / proviral DNA. The genetic characterization and phylogenetic analysis of FeLV and FIV were carried out in this study. These cats also tested for Leishmaniasis and Toxoplasmosis previously. FIV Ab and proviral DNA detected in 25.2 % and 25.5 % of samples, respectively. FeLV Ab, Ag, proviral DNA positivity was in 45.2 %, in 3.3 %, in 69.7 %, respectively. The molecular detection and phylo-genetic analysis of the current FeLV pol gene and FIV gag gene performed. The molecular characterization for the pol gene of FeLV (enFeLV and exFeLV) among Turkey's cat population was reported for the first time. The exFeLV pol sequences closer to the FeLV-A genotype, and the enFeLV pol sequences overlapped with other enFeLV. The current FIV gag sequences were clustered within the subtypes A, B, and C. The findings revealed FeLV subtype A and FIV subtype-A, subtype-B, subtype-C circulate among Turkish stray cats. Single and multiple co-infection positivity was found higher compared to previous reports.Öğe Molecular characterization of Anaplasma ovis Msp4 protein in strains isolated from ticks in Turkey: A multi-epitope synthetic vaccine antigen design against Anaplasma ovis using immunoinformatic tools(Academic Press Ltd- Elsevier Science Ltd, 2024) Koeseoglu, Ahmet Efe; Can, Hueseyin; Guvendi, Mervenur; Alak, Sedef Erkunt; Doskaya, Aysu Degirmenci; Karakavuk, Muhammet; Doskaya, MertTick-borne pathogens increasingly threaten animal and human health as well as cause great economic loss in the livestock industry. Among these pathogens, Anaplasma ovis causing a decrease in meat and milk yield is frequently detected in sheep in many countries including Turkey. This study aimed to reveal potential vaccine candidate epitopes in Msp4 protein using sequence data from Anaplasma ovis isolates and then to design a multiepitope protein to be used in vaccine formulations against Anaplasma ovis. For this purpose, Msp4 gene was sequenced from Anaplasma ovis isolates (n:6) detected in ticks collected from sheep in Turkey and the sequence data was compared with previous sequences from different countries in order to detect the variations of Msp4 gene/protein. Potential vaccine candidate and diagnostic epitopes were predicted using various immunoinformatics tools. Among the discovered vaccine candidate epitopes, antigenic and conserved were selected, and then a multi-epitope protein was designed. The designed vaccine protein was tested for the assessment of TLR-2, IgG, and IFN-g responses by molecular docking and immune simulation analyses. Among the discovered epitopes, EVASEGSGVM and YQFTPEISLV epitopes with properties of high antigenicity, non-allergenicity, and non-toxicity were proposed to be used for Anaplasma ovis in further serodiagnostic and vaccine studies.Öğe Molecular investigation of Blastocystis sp. and its subtypes in cancer patients under chemotherapy in Aegean region, Turkey(Elsevier, 2022) Oner, Tuelay Oncue; Karabey, Mehmet; Can, Huseyin; Doskaya, Aysu Degirmenci; Karakavuk, Muhammet; Gul, Aytuel; Koseoglu, Ahmet EfeBlastocystis 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.Öğe Molecular prevalence and genetic diversity of Bartonella spp. in stray cats of Izmir, Turkey(Bmc, 2022) Koseoglu, Ahmet Efe; Can, Huseyin; Guvendi, Mervenur; Karakavuk, Muhammet; Manyatsi, Pumla; Alak, Sedef Erkunt; Doskaya, Aysu DegirmenciBackground Bartonella spp. are vector-borne pathogens that cause zoonotic infections in humans. One of the most well-known of these is cat-scratch disease caused by Bartonella henselae and Bartonella clarridgeiae, with cats being the major reservoir for these two bacteria. Izmir, Turkey is home to many stray cats, but their potential role as a reservoir for the transmission of Bartonella to humans has not been investigated yet. Therefore, the aim of this study was to investigate the prevalence of Bartonella species and their genetic diversity in stray cats living in Izmir. Methods Molecular prevalence of Bartonella spp. in stray cats (n = 1012) was investigated using a PCR method targeting the 16S-23S internal transcribed spacer gene (ITS), species identification was performed by sequencing and genetic diversity was evaluated by haplotype analysis. Results Analysis of the DNA extracted from 1012 blood samples collected from stray cats revealed that 122 samples were Bartonella-positive, which is a molecular prevalence of 12.05% (122/1012; 95% confidence interval [CI] 10.1-14.2%). Among the Bartonella-positive specimens, 100 (100/122; 81.96%) were successfully sequenced, and B. henselae (45/100; 45%), B. clarridgeiae (29/100; 29%) and Bartonella koehlerae (26/100; 26%) were identified by BLAST and phylogenetic analyses. High genetic diversity was detected in B. clarridgeiae with 19 haplotypes, followed by B. henselae (14 haplotypes) and B. koehlerae (8 haplotypes). Conclusions This comprehensive study analyzing a large number of samples collected from stray cats showed that Bartonella species are an important source of infection to humans living in Izmir. In addition, high genetic diversity was detected within each Bartonella species.Öğe Molecular prevalence and subtyping of Cryptosporidium spp. in fecal samples collected from stray cats in Izmir, Turkey(Bmc, 2022) Koseoglu, Ahmet Efe; Can, Huseyin; Karakavuk, Muhammet; Guvendi, Mervenur; Doskaya, Aysu Degirmenci; Manyatsi, Pumla Bhekiwe; Doskaya, MertBackground Cryptosporidium spp. are obligate intracellular apicomplexan parasites transmitted to humans and other animals by contaminated water, food, or direct contact. They mainly cause gastrointestinal symptoms, although subclinical infections are also common. Cats are primarily infected by host-adapted Cryptosporidium felis while C. parvum and C. muris have also been detected in some cases. In this study, the molecular prevalence of Cryptosporidium spp. was investigated by screening 399 fecal samples collected from stray cats using nested PCR targeting the 18S rRNA gene for the first time in Turkey. Additionally, Cryptosporidium PCR-positive samples were genotyped by nested PCR- restriction fragment length polymorphism (RFLP), and subsequently, amplicons of 18S SSU rRNA were sequenced. They were further subtyped by amplification and sequencing of the gp60 gene. Results Among fecal samples screened, 12 of them (3%) were found to be Cryptosporidium-positive, and according to RFLP and sequencing of 18S rRNA gene, all positive samples were identified as C. felis. Subtyping analyses at the gp60 gene showed that C. felis isolates belonged to the XIXa subtype family, which are closely related to human subtypes of the parasite. Conclusions The results of this study are important in terms of indicating the potential role of stray cats for transmission of Cryptosporidium spp. to humans or other animals. Also, the presence of XIXa, which is the dominant subtype family of C. felis in cats and humans was shown for the first time in stray cats of Izmir, Turkey.Öğe Molecular prevalence of Blastocystis sp. and subtype diversity in fecal samples collected from cattle in dairy farms in Turkey(Elsevier Sci Ltd, 2022) Oner, Tuelay Oncu; Karakavuk, Muhammet; Doskaya, Aysu Degirmenci; Guvendi, Mervenur; Guel, Aytul; Koseoglu, Ahmet Efe; Alak, Sedef ErkuntClose 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.Öğe Newly developed peptide-ELISA successfully detected anti-IgG antibodies against Maedi-Visna virus in sheep(Elsevier, 2024) Kockaya, Ecem Su; Can, Huseyin; Yaman, Yalcin; Kandemir, Cagri; Taskin, Turgay; Karakavuk, Muhammet; Doskaya, Aysu DegirmenciMaedi Visna Virus (MVV) is a retrovirus that can infect sheep. There is still no effective therapy or vaccine against this virus and timely diagnosis is important to combat the complications of the disease. In this study, we aimed to develop an ELISA using peptides derived from gag protein as antigen. For this purpose, B cell epitopes of gag protein were predicted and a docking analysis with the B cell receptor was performed to select peptides to be used in ELISA. After three soluble epitopes with the highest antigenicity were produced as peptides, the immunogenicity of each peptide was determined by ELISA using sheep serum samples categorized as MVV positive (n=24) and negative (n=13). Subsequently, in house ELISA using above mentioned immunogenic peptides as antigen was used to investigate MVV seroprevalence in sheep (n=88). According to the results, among three peptides, two of them strongly reacted with MVV positive serum samples and the mean absorbance values detected among positive and negative serum samples were statistically significant, indicating that these peptides were immunogenic (P=0.016 and P=0.038). The third peptide also reacted with positive serum samples but the mean absorbance value was not statistically significant and this peptide was considered non-immunogenic (P=0.175). The immunogenic two peptides showed the same high sensitivity and specificity values of 91.60 and 92.80 according to the commercial kit. Moreover, MVV seroprevalence detected by peptide-ELISAs using CKQGSKE and CRPQGKAGHKG peptides as antigen was 3.40 % and 4.5 %, respectively. As a result, it was shown that these peptides can be successfully used for serological diagnosis of MVV.Öğe Preliminary analysis of Plasmodium vivax genotypes isolated in southeastern Turkey(Walter De Gruyter Gmbh, 2015) Doskaya, Aysu Degirmenci; Doskaya, Mert; Caner, Ayse; Gul, Kadri; Nergiz, Sebnem; Can, Huseyin; Guruz, YukselPlasmodium vivax is the most common cause of malaria worldwide as well as southeastern Turkey. After the implementation of a successful national elimination program that the local malaria cases were not reported in 2011, malaria returned to county of Savur located in southeastern Turkey in summer of 2012. The present study aimed to determine the prevalent P. vivax genotypes isolated from southeastern Turkey. Genetic polymorphism in P. vivax CSP gene was analyzed by PCR-RFLP to assess the ratio of VK210 and VK247 types. Blood samples were obtained from 15 patients who lived in southeastern between 2005-2006. According to the results, VK210 type was detected in 10 samples (66.6%), VK247 type was observed in three samples (20%). Remaining two samples showed mixed infection (13.3%). The results of the present study first time showed the ratio of P. vivax genotypes in southeastern Turkey before the elimination in 2011. The results of the present study will be enable researchers to compare the new isolates with the previously detected ones and design new treatment and/elimination strategies.Öğe Preliminary analysis of Plasmodium vivax genotypes isolated in southeastern Turkey(Walter De Gruyter Gmbh, 2015) Doskaya, Aysu Degirmenci; Doskaya, Mert; Caner, Ayse; Gul, Kadri; Nergiz, Sebnem; Can, Huseyin; Guruz, YukselPlasmodium vivax is the most common cause of malaria worldwide as well as southeastern Turkey. After the implementation of a successful national elimination program that the local malaria cases were not reported in 2011, malaria returned to county of Savur located in southeastern Turkey in summer of 2012. The present study aimed to determine the prevalent P. vivax genotypes isolated from southeastern Turkey. Genetic polymorphism in P. vivax CSP gene was analyzed by PCR-RFLP to assess the ratio of VK210 and VK247 types. Blood samples were obtained from 15 patients who lived in southeastern between 2005-2006. According to the results, VK210 type was detected in 10 samples (66.6%), VK247 type was observed in three samples (20%). Remaining two samples showed mixed infection (13.3%). The results of the present study first time showed the ratio of P. vivax genotypes in southeastern Turkey before the elimination in 2011. The results of the present study will be enable researchers to compare the new isolates with the previously detected ones and design new treatment and/elimination strategies.Öğe Preliminary analysis of Plasmodium vivax genotypes isolated in southeastern Turkey(Walter De Gruyter Gmbh, 2015) Doskaya, Aysu Degirmenci; Doskaya, Mert; Caner, Ayse; Gul, Kadri; Nergiz, Sebnem; Can, Huseyin; Guruz, YukselPlasmodium vivax is the most common cause of malaria worldwide as well as southeastern Turkey. After the implementation of a successful national elimination program that the local malaria cases were not reported in 2011, malaria returned to county of Savur located in southeastern Turkey in summer of 2012. The present study aimed to determine the prevalent P. vivax genotypes isolated from southeastern Turkey. Genetic polymorphism in P. vivax CSP gene was analyzed by PCR-RFLP to assess the ratio of VK210 and VK247 types. Blood samples were obtained from 15 patients who lived in southeastern between 2005-2006. According to the results, VK210 type was detected in 10 samples (66.6%), VK247 type was observed in three samples (20%). Remaining two samples showed mixed infection (13.3%). The results of the present study first time showed the ratio of P. vivax genotypes in southeastern Turkey before the elimination in 2011. The results of the present study will be enable researchers to compare the new isolates with the previously detected ones and design new treatment and/elimination strategies.
