The molecular and serological investigation of Feline immunodeficiency virus and Feline leukemia virus in stray cats of Western 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 phylogenetic 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. © 2021 Elsevier Lt

Analysis of cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene related to neonatal isoerythrolysis in stray cats of Izmir, Turkey [Türkiye, İzmir sokak kedilerinde neonatal izoeritrolizisle ilişkili sitidin monofosfat-N-asetilnöraminik asit hidroksilaz (CMAH) geninin analizi]

Neonatal isoerythrolysis is a life threatening disease in new born cats. It occurs when type A or type AB kittens are born from a type B queen (female cat). A homozygous 18 bp insertion located in cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) gene has been detected in type B cats, causing production of inactive CMAH enzyme. Currently, molecular methods are being used to determine type B blood in female cats, which can help prevent neonatal isoerythrolysis in kittens. These molecular assays target the presence of 18 bp insertion in CMAH gene. In this study, we aimed to analyze the potential of neonatal isoerythrolysis among stray cats of İzmir, Turkey using PCR detecting the 18 bp insertion in CMAH gene. During the study, we analyzed 793 cats’ blood sample for the presence of 18 bp insertion in CMAH gene. Three cats known to have blood types A, B, and AB were used as control in PCR. According to the PCR results, blood type A control cat displayed a 175 bp product indicating a homozygous type A cat while blood type control B cat showed a 193 bp product in CMAH gene (with 18 bp insertion) indicating a homozygous type B cat. Interestingly, blood type AB control cat showed a heterozygous pattern for CMAH gene, in which three different bands (175 bp like that of type A, 193 bp product for type B, and the third unique band with approximately 240 bp size) were detected. Among 793 stray cats of İzmir, 791 were homozygous for CMAH gene with 175 bp band size (99.7%). The remaining two stray cats showed heterozygous band pattern like blood type AB cat (0.12%). Overall, 175 bp band displaying type A cats are prevalent contrary to the two cats that have type AB pattern and non-existence of homozygous type B cats. These results show that the potential of neonatal isoerythrolysis in stray cats of İzmir is minimal. Future studies are required to scrutinize the reason(s) for non-existence of type B cats in İzmir and presence of unique band in blood type AB. © 2016, Veteriner Fakultesi Dergisi. All rights reserved

Genotyping of Pneumocystis jirovecii isolates obtained from clinical samples by multilocus sequencing: a molecular epidemiology study conducted in Turkey

PubMed: 322745572-s2.0-85083059533Pneumocystis jirovecii is an opportunistic respiratory pathogen causing Pneumocystis pneumonia (PcP) in immunocompromised patients. The aim of this study was to investigate the genetic diversity of P. jirovecii isolates (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 for P. jirovecii DNA, 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. © 2020, Springer-Verlag GmbH Germany, part of Springer Nature.2013TIP050, TGA-2019-20253This study was supported by Ege University Scientific Research Projects Coordination Unit (Project Numbers: TGA-2019-20253 and 2013TIP050)

[Demonstration of Cryptosporidium parvum in immune suppressed rats using nested PCR] [İmmun Sistemi Baskılanmış Sıçanlarda Cryptosporidium parvum'un Nested PZR Yöntemi ile Gösterilmesi]

PubMed ID: 24192616[No abstract available

A preliminary study to develop a lateral flow assay using recombinant GRA1 protein for the diagnosis of toxoplasmosis in stray cats

Toxoplasma gondii is a protozoan parasite that may infect many mammals including humans. Cats are one of the main sources of infection for humans. Therefore, routine screening of cats with tests that are inexpensive, rapid, and do not require sophisticated laboratory equipment is important. In this study, a lateral flow assay (LFA) was designed to rapidly diagnose toxoplasmosis in cats. For this purpose, we selected GRA1 protein of T. gondii due to its high antigenicity in diagnostic and vaccine studies. We further analyzed the immunological properties of GRA1 protein using in silico tools. Then, we expressed and purified recombinant GRA1 (rGRA1) protein and used it during the development of LFA to detect toxoplasmosis in serum samples (n = 40) of cats. According to the results, rGRA1 protein has negative GRAVY value, high aliphatic index, alpha helix, random coil and 12 B cell epitopes. The in silico data supported the high antigenic properties of rGRA1 protein and showed that it can be a good antigen candidate for LFA. Among 30 cat positive serum samples, 27 were found positive by the LFA while seronegative sera (n = 10) were negative by the LFA. The preliminary data showed that the LFA has high sensitivity (90 %) and specificity (100 %). When we used high responsive cat sera (i.e. sera that have optical density > 0.5 with ELISA) the sensitivity value reached 100 %. These results showed that rGRA1 protein is a good candidate to develop a LFA for rapid diagnosis of toxoplasmosis in cats. © 2023 Elsevier LtdThis study was supported by a project given by the Ege University Scientific Research Projects Coordination Unit (Project no.: TGA-2021-22468 ) to A.D.D.Ege Üniversitesi: TGA-2021-2246