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Breast cancer was ranked first in global cancer incidence in 2020, and HER2 overexpression in breast cancer accounts for 20-30% of breast cancer patients. Current therapeutic strategies increase the survival rate, but resistance to them occurs frequently, and there is an urgent need to develop novel treatments such as DNA vaccines which can induce a specific and long-lasting immune response against HER2 antigens. To enhance the immunogenicity of DNA vaccines, dendritic cells (DCs) can be targeted using multiepitope proteins that provide accurate immune focusing. For this purpose, we generated a DNA vaccine encoding a fusion protein composed of 1) in silico discovered antigenic epitopes of human and rat HER2 proteins (MeHer2) and 2) a single-chain antibody fragment (ScFv) specific for the DC-restricted antigen uptake receptor DEC205 (ScFvDEC). The xenogeneic multi-epitope DNA vaccine (pMeHer2) encodes three only T-cell epitopes, two only B-cell epitopes, and two T and B cell epitopes, and pScFvDEC-MeHer2 vaccine additionally encodes ScFvDEC introduced at the N terminus of the MeHer2. Then, mouse groups were immunized with pScFvDEC-MeHer2, pMeHer2, pScFvDEC, pEmpty, and PBS to determine the elicited immune response. pScFvDEC-MeHer2 vaccinated mice showed a strong IgG response (P < 0.0001) and pScFvDEC-MeHer2 induced a significant IgG2a increase (P < 0.01). The percentages of both IFN-gamma secreting CD4 and CD8 T cells were higher in mice immunized with pScFvDEC-MeHer2 compared with the pMeHer2. pScFvDEC-MeHer2 and pMeHer2 secreted significantly higher levels of extracellular IFN-gamma compared with to control groups (P < 0.0001). In addition, the IFN-gamma level of the pScFvDEC-MeHer2 vaccine group was approximately two times higher than the pMeHer2 group (P < 0.0001). Overall, this study identified the pScFvDECMeHer2 construct as a potential DNA vaccine candidate, supporting further studies to be conducted on HER2(+)& nbsp;animal models.(c) 2022 Elsevier Ltd. All rights reserved., Scientific Research Projects Branch Directorate of Ege University, Turkey [2016-TIP-082], Acknowledgement This study was supported by the grant given by the Scientific Research Projects Branch Directorate of Ege University, Turkey (Grant No: 2016-TIP-082) to L.Y.

Ehrlichia spp. is tick-borne zoonotic pathogen that can infect humans and animals. Nowadays, among the tests used in the diagnosis of ehrlichiosis, the importance of molecular methods is increasing steadily due to their high sensitivity and specificity. The aim of this study was to determine the analytical sensitivity of a conventional polymerase chain reaction (PCR) targeting Ehrlichia spp. disulfide oxidoreductase (DSB) gene. Ehrlichia spp. DSB gene was cloned into the TOPO vector. After TOPO plasmid containing DSB gene were serially diluted, PCR targeting the Ehrlichia spp. DSB gene was performed. While working on this research, blood and skin scraping samples of a stray dog clinically suspected with leishmaniasis as well as treated for leishmaniasis arrived to our laboratory. Thereafter, PCRs targeting Ehrlichia spp. DSB and 16S rRNA and Leishmania kinetoplast DNA (kDNA) genes were performed to identify the pathogen in blood and skin scraping samples of the stray dog. The analytical sensitivity of the PCR assay targeting Ehrlichia spp. DSB gene was 1 ≥ copy plasmid/reaction using serially diluted TOPO plasmid containing DSB gene. PCR targeting the Ehrlichia spp. DSB gene was positive and PCR targeting Leishmania spp. kDNA was negative in blood and skin samples of the stray dog clinically suspected with leishmaniasis. Using nested PCR targeting Ehrlichia spp. 16S rRNA, E. canis was identified in blood and skin scraping samples of the stray dog. In this study, PCR targeting Ehrlichia spp. DSB gene has been shown to have high sensitivity. Also it was shown molecular methods can help clinicians in differential diagnosis of ehrlichiosis and leishmaniasis to prevent inappropriate treatment.

Amaç: Toxoplasma gondii insan ve sıcakkanlı hayvanlarda hastalıklara neden olan bir protozoondur. Bu çalışmanın amacı toksoplazmozis tanısı için T. gondii tekrar bölgesine (RE) özgü primerlerin tasarlanması ile geliştirilen floresans esaslı hızlı Döngü Aracılı İzotermal Amplifikasyon (LAMP) testinin analitik hassasiyetini belirlemektir.Gereç ve Yöntem: Polimeraz Zincir Reaksiyonu (PZR) ile elde edilen T. gondii RE, pCR2.1-TOPO vektörüne klonlanmıştır. Spesifik primerler kullanılarak geliştirilen LAMP testinin analitik hassasiyeti pCR2.1-RE vektörünün seri seyreltmeleri ile belirlenmiştir.Bulgular: T. gondii RE genine özgü tasarlanan primerler ile geliştirilen LAMP testinin analitik hassasiyetinin ≤10 plazmit/reaksiyon olduğu tespit edilmiştir.Sonuç: RE bölgesi T. gondii geni içinde 200-300 kere tekrar ettiğinden dolayı geliştirilen testin 0,05 takizoit/reaksiyon saptama limitine sahip olduğu hesaplanmıştır. LAMP testinin kolay, ucuz, hızlı ve saha koşullarına uygun olması ve diğer moleküler testlerden daha hassas olması nedeniyle umut verici olduğu öngörülmektedir, Aim: Toxoplasma gondii is a protozoon that causes disease in humans and warm-blooded animals. The aim of this study is to determine the analytical sensitivity of the fluorescence-based rapid loop mediated isothermal amplification (LAMP) test developed by designing primers specific to T. gondii repeat region (RE) for the diagnosis of toxoplasmosis.Materials and Methods: T. gondii RE obtained with PCR was cloned into vector pCR2.1-TOPO. The analytical sensitivity of the LAMP assay developed using specific primers was determined by serial dilutions of the pCR2.1-RE vectorResults: It was determined that the analytical sensitivity of the LAMP test developed with primers designed specific to T. gondii RE gene was 10 copy plasmid / reaction.Conclusion: Since the RE region repeats 200-300 times in the T. gondii gene, the test developed has been calculated to have a detection limit of 0.05 tachyzoites/reaction. The LAMP test is predicted to be promising because it is easy, cheap, fast and suitable for field conditions and is more sensitive than other molecular tests

Background: Tick-borne pathogens increasingly threaten animal and human health as well as cause great economic loss in the livestock industry. Among these pathogens, Anaplasma oviscausing a decrease in meat and milk yield is frequently detected in sheep in many countries including Turkey. Unfortunately a vaccine has not been developed against Anaplasma ovis yet. Msp4 is a immunogenic surface protein of Anaplasma bacteria. This study aimed to reveal potential vaccine candidate epitopes in Msp4 protein using sequence data belonging to Msp4 gene of Anaplasma ovis isolates and then to design a multi-epitope protein to be used in vaccine formulations against Anaplasma ovis. Results: 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. Later, potential vaccine candidate and diagnostic epitopes were predicted using various immunoinformatics tools including physicochemical parameters, secondary structures, antigenicity, solubility, subcellular localization, transmembrane helix, signal peptide, allergenicity, BetaWrap motif, homology checking, postranslational modifications, B/T cell epitopes, toxicity, solvent exposure, and epitope conservancy. 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. Finally, among the discovered epitopes of Anaplasma ovis, EVASEGSGVM and YQFTPEISLV epitopes that have properties of high antigenicity, non-allergenicity, and non-toxicity were proposed for the development of peptide-based vaccines and serodiagnostic assays. Conclusions: This study provides prominent epitopes and a candidate vaccine model to be used for the tick-borne pathogen Anaplasma ovis in further serodiagnostic and vaccine studies.

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral infection caused by Crimean-Congo hemorrhagic fever virus (CCHFV). Serological screening of CCHF is important and current ELISA use antigens prepared from virus which is expensive due to requirement of high bio-containment facilities. in this study, we aimed to develop a new recombinant ELISA. For this purpose, CCHFV genome were expressed as 13 proteins in E. coli and among them abundantly purified recombinant Nucleocapsid protein (rNP) and Mucin-like variable domain (rMLD) were used as antigen in ELISA (Rec-ELISA). Rec-ELISA using rNP, rMLD and a combination of both (rNP/rMLD) were probed with acute (n=64; collected between days 1 and 7 after onset of symptoms), convalescent (n=35; collected 8 days after onset of symptoms), consecutive sera (n=25) of confirmed CCHF cases and control sera (n=43). The sensitivity and specificity of Rec-ELISA using rNP/rMLD were 73% and 98% in acute cases and 97% and 98% in convalescent cases. The median interquartile absorbance value to discriminate the acute and convalescent phases of CCHF was significantly higher with ELISA using rNP/rMLD (P0.05) and rMLD (P=0.001). These results indicate that the Rec-ELISA using rNP/rMLD may be very useful to diagnose convalescent CCHF cases especially in field studies., Turkish Scientific and Technological Research Council (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK), The authors would like to acknowledge Turkish Scientific and Technological Research Council (TUBITAK) for supporting Sultan Gulce-z by a 2219 post-doctoral research fellowship programme to conduct part of the studies at Department of Infectious Diseases, University of California, Irvine.

Introduction: 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. © 2021 Journal of Infection in Developing Countries. All rights reserved., 2016-TIP-042, This study was supported by the grant given by the Scientific Research Projects Branch Directorate of Ege University, Turkey (Grant No: 2016-TIP-042) to M.D.

Toxoplasma gondii is a zoonotic protozoan parasite that can infect humans and warm-blooded animals. In this study, it was aimed to investigate the frequency of toxoplasmosis in clinical samples taken from various patient groups. Within the scope of the study, a total of 535 clinical samples, including mainly amniotic fluid, blood and cerebrospinal fluid samples, which were sent to the Molecular Parasitology laboratory of Ege University Faculty of Medicine, Department of Parasitology between 2009 and 2019, were evaluated.The frequency of toxoplasmosis was determined as 2.61% (14/535) in the specified time period. While the positivity rate was 5.40% (8/148) in the samples taken from the central organs directly affected by the disease, the positivity rate was 1.74% (5/286) in the samples taken from the periphery, and this difference was statistically significant (P0.05).The findings show that Real-Time PCR method has a very important place in the diagnosis of active toxoplasmosis. In addition, it was concluded that sampling methods are very important in the diagnosis of the disease., Toxoplasma gondii insan ve sıcakkanlı hayvanları enfekte edebilen zoonotik protozoon bir parazittir. Bu çalışmada çeşitli hasta gruplarından alınmış klinik örneklerde toksoplazmozis sıklığının araştırılması amaçlanmıştır. Çalışma kapsamında 2009-2019 yılları arasında Ege Üniversitesi Tıp Fakültesi Parazitoloji Anabilim Dalı Moleküler Parazitoloji laboratuvarına gönderilen ağırlıklı olarak amniyon sıvısı, kan ve beyin omurilik sıvısı örneklerinin dahil olduğu toplam 535 klinik örnek değerlendirilmiştir. Bu örneklerde DNA izolasyonu sonrası T. gondii RE geni varlığı Real-Time PZR ile araştırılmıştır.Belirtilen zaman aralığında toksoplazmozis sıklığı %2,61 (14/535) olarak tespit edilmiştir. Hastalığın direktetkilediği merkezi organlardan alınan örneklerde pozitiflik oranı %5,40 (8/148) iken periferden alınan örneklerde pozitiflik oranı %1,74 (5/286) olarak tespit edilmiş ve bu fark istatistiksel olarak anlamlı bulunmuştur (P0,05).Elde edilen bulgular toksoplazmozis tanısında Real-Time PZR yönteminin oldukça önemli bir yeri olduğunugöstermektedir. Ayrıca, örnek alım yöntemlerinin hastalığın tanısında oldukça önemli olduğu sonucuna varılmıştır.

Toxoplasma gondii insan ve sıcakkanlı hayvanları enfekte edebilen zoonotik protozoon bir parazittir. Bu çalışmada çeşitli hasta gruplarından alınmış klinik örneklerde toksoplazmozis sıklığının araştırılması amaçlanmıştır. Çalışma kapsamında 2009-2019 yılları arasında Ege Üniversitesi Tıp Fakültesi Parazitoloji Anabilim Dalı Moleküler Parazitoloji laboratuvarına gönderilen ağırlıklı olarak amniyon sıvısı, kan ve beyin omurilik sıvısı örneklerinin dahil olduğu toplam 535 klinik örnek değerlendirilmiştir. Bu örneklerde DNA izolasyonu sonrası T. gondii RE geni varlığı Real-Time PZR ile araştırılmıştır. Belirtilen zaman aralığında toksoplazmozis sıklığı %2,61 (14/535) olarak tespit edilmiştir. Hastalığın direkt etkilediği merkezi organlardan alınan örneklerde pozitiflik oranı %5,40 (8/148) iken periferden alınan örneklerde pozitiflik oranı %1,74 (5/286) olarak tespit edilmiş ve bu fark istatistiksel olarak anlamlı bulunmuştur (P0,05). Elde edilen bulgular toksoplazmozis tanısında Real-Time PZR yönteminin oldukça önemli bir yeri olduğunu göstermektedir. Ayrıca, örnek alım yöntemlerinin hastalığın tanısında oldukça önemli olduğu sonucuna varılmıştır.

Breast cancer was ranked first in global cancer incidence in 2020, and HER2 overexpression in breast cancer accounts for 20-30% of breast cancer patients. Current therapeutic strategies increase the survival rate, but resistance to them occurs frequently, and there is an urgent need to develop novel treatments such as DNA vaccines which can induce a specific and long-lasting immune response against HER2 antigens. To enhance the immunogenicity of DNA vaccines, dendritic cells (DCs) can be targeted using multi-epitope proteins that provide accurate immune focusing. For this purpose, we generated a DNA vaccine encoding a fusion protein composed of 1) in silico discovered antigenic epitopes of human and rat HER2 proteins (MeHer2) and 2) a single-chain antibody fragment (ScFv) specific for the DC-restricted antigen-uptake receptor DEC205 (ScFvDEC). The xenogeneic multi-epitope DNA vaccine (pMeHer2) encodes three only T-cell epitopes, two only B-cell epitopes, and two T and B cell epitopes, and pScFvDEC-MeHer2 vaccine additionally encodes ScFvDEC introduced at the N terminus of the MeHer2. Then, mouse groups were immunized with pScFvDEC-MeHer2, pMeHer2, pScFvDEC, pEmpty, and PBS to determine the elicited immune response. pScFvDEC-MeHer2 vaccinated mice showed a strong IgG response (P 0.0001) and pScFvDEC-MeHer2 induced a significant IgG2a increase (P 0.01). The percentages of both IFN-γ secreting CD4 and CD8 T cells were higher in mice immunized with pScFvDEC-MeHer2 compared with the pMeHer2. pScFvDEC-MeHer2 and pMeHer2 secreted significantly higher levels of extracellular IFN-γ compared with to control groups (P 0.0001). In addition, the IFN-γ level of the pScFvDEC-MeHer2 vaccine group was approximately two times higher than the pMeHer2 group (P 0.0001). Overall, this study identified the pScFvDECMeHer2 construct as a potential DNA vaccine candidate, supporting further studies to be conducted on HER2

The cat flea Ctenocephalides felis has veterinary and medical importance since it is a vector for numerous important pathogens. In this study, a total of 249 flea samples were collected from goats bred in eight different farms (located in Izmir and S,anliurfa provinces of Turkey) and morphologically identified under microscopy. Later, the genetic diversity was investigated in 117 of C. felis samples that were morphologically identified by sequencing the mitochondrial cox1 gene, followed by phylogenetic tree, haplotype, genetic differentiation and gene flow analyses. In addition, Rickettsia spp. and Bartonella spp. which are zoonoses were screened in 27 pools comprising 249 flea samples by PCR. The phylogenetic tree showed that 117 flea samples were clustered in Clade 1 together with isolates from Australia, New Zealand, the Czech Republic, and India. Four haplotypes (haplo-types I, II, III and IV) were detected within the C. felis species. The most prevalent haplotype was haplotype I (57/ 117; 48.7 %). Among the population of flea samples in Izmir and S,anliurfa, the Fst and Nm values were 0.16261 and 2.57, respectively, indicating a moderate genetic differentiation and high gene flow. Rickettsia spp. was detected in four of C. felis pool samples whereas Bartonella spp. was detected in 25 of them. BLAST analysis identified R. raoultii as well as B. henselae and B. elizabethae. In conclusion, the findings showed that C. felis samples collected from goats in Turkey were classified within Clade 1 representing four different haplotypes with a moderate genetic diversity for the first time. Also, R. raoultii, B. henselae and B. elizabethae were demonstrated for the first time in cat flea samples collected in Turkey., Ege University Scientific Research Projects Coordination Unit; [TGA- 2019-20233], Funding This study was supported by a project given by the Ege University Scientific Research Projects Coordination Unit (Project number: TGA- 2019-20233) to H.C.

Blastocystis 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., Scientific and Technological Research Council of Turkey (TUEB ?ITAK) [119S785]; Izmir Katip Celebi University Scientific Research Projects Coordination Unit [2018-TDU-TIPF-0055], This study was partially supported by a project provided by The Scientific and Technological Research Council of Turkey (TUEB ?ITAK) (Project number: 119S785) to H.C and Izmir Katip Celebi University Scientific Research Projects Coordination Unit (Project number: 2018-TDU-TIPF-0055) to A.A.G.

Close 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, Scientific and Technological Research Council of Turkey (TUBITAK) [119S785]; Ege University Scientific Research Projects Coordination Unit [TGA-020-20850], This study was partially supported by a project provided by The Scientific and Technological Research Council of Turkey (TUBITAK) (Project number: 119S785) to H.C and Ege University Scientific Research Projects Coordination Unit (Project number: TGA-020-20850) to M.K.

BackgroundThe emergence of tick-borne disease is increasing because of the effects of the temperature rise driven by global warming. In Turkey, 19 pathogens transmitted by ticks to humans and animals have been reported. Based on this, this study aimed to investigate tick-borne pathogens including Hepatozoon spp., Theileria spp., Babesia spp., Anaplasma spp., Borrelia spp., and Bartonella spp. in tick samples (n = 110) collected from different hosts (dogs, cats, cattle, goats, sheep, and turtles) by molecular methods.MethodsTo meet this objective, ticks were identified morphologically at the genus level by microscopy; after DNA isolation, each tick sample was identified at the species level using the molecular method. Involved pathogens were then investigated by PCR method.ResultsSeven different tick species were identified including Rhipicephalus sanguineus, R. turanicus, R. bursa, Hyalomma marginatum, H. anatolicum, H. aegyptium, and Haemaphysalis erinacei. Among the analyzed ticks, Hepatozoon spp., Theileria spp., Babesia spp., and Anaplasma spp. were detected at rates of 6.36%, 16.3%, 1.81%, and 6.36%, respectively while Borrelia spp. and Bartonella spp. were not detected. Hepatozoon spp. was detected in R. sanguineus ticks while Theileria spp., Babesia spp., and Anaplasma spp. were detected in R. turanicus and H. marginatum. According to the results of sequence analyses applied for pathogen positive samples, Hepatozoon canis, Theileria ovis, Babesia caballi, and Anaplasma ovis were identified.ConclusionTheileria ovis and Anaplasma ovis were detected for the first time to our knowledge in H. marginatum and R. turanicus collected from Turkey, respectively. Also, B. caballi was detected for the first time to our knowledge in ticks in Turkey., Ege University Scientific Research Projects Coordination Unit [TGA-2020-21040], This study was supported by a project given by the Ege University Scientific Research Projects Coordination Unit (Project number: TGA-2020-21040) to H.C.

Toxoplasma 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., Scientific and Technological Research Council of Turkey [119S367], This study was supported by the Grant given by The Scientific and Technological Research Council of Turkey (119S367) to M.D. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Amaç: Toksoplasma gondii insan ve sıcakkanlı hayvanlarda hastalıklara neden olan bir protozoondur. Bu çalışmanın amacı toksoplazmozis tanısı için T.gondii tekrar bölgesine (RE) özgü primerlerin tasarlanması ile geliştirilen floresans esaslı hızlı LAMP testinin analitik hassasiyetini belirlemektir. Gereç ve Yöntem: PZR ile elde edilen T. gondii RE, pCR2.1-TOPO vektörüne klonlanmıştır. Spesifik primerler kullanılarak geliştirilen LAMP testinin analitik hassasiyeti pCR2.1-RE vektörünün seri seyreltmeleri ile belirlenmiştir. Bulgular: T. gondii RE genine özgü tasarlanan primerler ile geliştirilen LAMP testinin analitik hassasiyetinin 10≤kopya plazmit/reaksiyon olduğu tespit edilmiştir. Sonuç: Geliştirilen test 0,05 takizoit saptama limitine sahip olup oldukça hassas bir testtir. LAMP testinin kolay, ucuz, hızlı ve saha koşullarına uygun olması ve diğer moleküler testlerden daha hassas olması nedeniyle umut verici olduğu öngörülmektedir.

Gastrointestinal parasites of cats can affect animal health and welfare, as well as human health because of some zoonotic parasites including Toxoplasma gondii, Cryptosporidium spp., Isospora spp., Blastocystis sp., and Toxocara spp. Therefore, it is fairly important to investigate the presence of gastrointestinal parasites in stray cats in order to reveal the frequency of parasite diseases and to prevent the spread of parasitic diseases. A total of 465 feces samples were collected from Veterinary Clinics located in 5 different districts of İzmir. For microscopic examination, all feces samples were processed by centrifugation-sucrose flotation. In addition, cat feces with diarrhea were stained by the by Kinyoun acid-fast staining for the diagnosis of Cryptosporidium spp. As a result, 73 of 465 (15.6%) cats were found to be infected with at least one of the following parasites: Blastocystis sp., Isospora spp., Cryptosporidium spp., Toxoplasma gondii-like oocyte, Toxocara spp., Hymenolepis spp. and Dipylidium caninum. Among the studied stray cats, Blastocystis sp. was detected as the most prevalent protozoon parasite (10.5%) in stray cats. Overall, the results show that stray cats are a significant source for distribution of various parasite diseases to humans and animals in İzmir, Turkey.

This 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.