Türkiye'deki Vahşi Kuşlarda Neospora caninum DNA'sının Varlığı

OBJECTIVE: Neospora caninum is a protozoon parasite that has a worldwide distribution and mainly causes abortion in cattle and current serological evidence shows that the disease may be also zoonotic. Wild birds play a role as a reservoir of the disease in nature. The study aimed to determine the prensence of N. caninum in wild birds. METHODS: In this study, the presence of neosporosis in wild birds (n=55) including 22 different species found in the western side of Turkey, was investigated by polymerase chain reaction (PCR). In addition, PCR positive samples were confirmed by sequencing, BLAST, and phylogenetic analysis using MEGA7. RESULTS: Obtained results showed that the presence of N. caninum DNA was 5.45% (3/55) in brain-heart homogenates wild birds. The bird species which were found positive for N. caninum were little owl (Athene noctua), common buzzard (Buteo buteo), and little tern (Sternula albifrons). According to phylogenetic analysis and BLAST, all samples were compatible with reference N. caninum isolates. CONCLUSION: To the best of authors' knowledge, this is the first study detecting N. caninum in little tern. In future studies, it may be interesting to investigate the prevalence of N. caninum in other wild animals to elucidate the transmission properties.Amaç: Neospora caninum, dünya çapında dağılım gösteren ve esas olarak sığırlarda düşüklere neden olan protozoon bir parazittir ve güncel serolojik kanıtlar hastalığın zoonotik olabileceğini göstermektedir. Yabani kuşlar, doğada hastalığın rezervuarı olarak rol oynamaktadır. Çalışma, yabani kuşlarda N. caninum varlığının belirlenmesini amaçladı. Yöntemler: Bu çalışmada, Türkiye’nin batısındaki 22 farklı yabani kuşta (n=55) neosporosis varlığı polimeraz zincir reaksiyon (PZR) ile araştırılmıştır. Ek olarak, PZR pozitif örnekler sekanslanarak MEGA7 kullanılarak BLAST ve filogenetik analiz ile doğrulanmıştır. Bulgular: Elde edilen sonuçlara göre, yabani kuşların beyin-kalp homojenatlarının %5,45’inde (3/55) N. caninum DNA’sı saptanmıştır. Kukumav (Athene noctua), bayağı şahin (Buteo buteo) ve küçük sumru (Sternula albifrons) N. caninum pozitif bulunan kuş türleridir. Filogenetik analiz ve BLAST sonuçlarına göre, tüm örnekler referans N. caninum izolatları ile uyumlu olarak tespit edilmiştir. Sonuç: Yazarların bildiği kadarıyla bu çalışma, küçük sumruda N. caninum tespit eden ilk çalışmadır. Gelecekteki çalışmalarda N. caninum bulaşma özelliklerini aydınlatmak amacıyla diğer vahşi hayvanlarda prevalansın araştırılması faydalı olabilir. Anahtar Kelimeler: Küçük sumru, Neospora caninum, PZR, Türkiye, vahşi kuşlar

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

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

PubMed ID: 25268351Toxoplasma 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. Copyright: © 2014 Döşkaya et al

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)

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