Browsing by Author "Akar, Kadir"

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Comparative Assessment of Brucellosis Detection in Dogs: In-House Elisa Versus Rose Bengal Plate Test Utilizing Rough and Smooth Antigens
(Elsevier Sci Ltd, 2025) Akar, Kadir; Yuecetepe, Ayfer Guellue; Ekin, Ismail Hakki; Dadar, Maryam; Guerbilek, Sevil Erdenlig
Brucella canis and smooth Brucella species infections have been reported in dogs globally. In endemic countries such as T & uuml;rkiye, dogs can be infected with both species. The exact incidence of canine brucellosis in T & uuml;rkiye is unknown. This study aims to investigate the seroprevalence of brucellosis in stray dogs undergoing sterilization in Van province, eastern T & uuml;rkiye. In this study, a comprehensive approach was used to ensure reliable incidence of Brucella spp in the dogs. We used the Rose Bengal Plate Test (RBPT) with both antigens from smooth Brucella abortus S99 (RBPT-S) and rough B. canis (RBPT-R) Brucella species to analyze 150 dog blood serum samples. For the in-house enzyme-linked immunosorbent assay (i-ELISA), we used a rough Lipopolysaccharide (LPS) antigen from the rough strain (i-ELISA-R) and an O-polysaccharide (O-PS) antigen (i-ELISA-S) from the smooth strain obtained from the OIE Reference Laboratory in Weybridge, UK. Each serum sample underwent analysis using a total of four serological tests, ensuring a thorough and reliable evaluation. Out of 150 serum samples analyzed, 32 (21.3 %) tested positive using the RBPT-S test and 12 (6.6 %) with the RBPT-R test. The i-ELISA-R identified 8 (5.3 %) positive samples, while the i-ELISA-S detected 27 (18 %) positive samples. The presence of B. canis and other Brucella spp. infections was detected in the dog in this region, revealing that canine brucellosis was primarily caused by smooth Brucella species. The test beased -smooth antigens identified a significantly higher number of positive samples compared to the test beased -rough antigens test, suggesting a potential difference in sensitivity or specificity between the two methods. This situation was considered an important risk for both dogs and humans, underscoring the urgent need for further research and the immediate implementation of public health measures to address this issue.
Development of a Simple and Rapid Dna Extraction Method for Aspergillus Flavus
(Sciendo, 2024) Golen, Gokcenur Sanioglu; Akar, Kadir
Aspergillus species are known to be very important in human and domestic animal health. Aspergillus species commonly cause severe systemic and skin infections, as well as allergic lung diseases. With the development of PCR techniques, these methods are used to identify and diagnose fungi. DNA extraction from Aspergillus species is difficult because the fungal cell wall structure is very durable and complex. Fungal DNA extraction methods containing proteinase K and liquid nitrogen are widely used to break down the cell wall. However, these methods cause DNA loss during the extraction in Aspergillus species. In this study, on the contrary, the commonly used DNA extraction by means of ammonium hydroxide, which is generally used to break down chitin in DNA extraction of ticks and plants, is used. The efficiency of the cell wall lysis method from A. flavus with ammonium hydroxide was compared with methods containing proteinase K and liquid nitrogen. For this purpose, DNA extraction of A. flavus was tried using three different methods. As a result, the cell wall of A. flavus was lysed using ammonium hydroxide in this study. The obtained DNA's quality, concentration, and PCR performance were sufficient. This method has been evaluated as a faster, more straightforward, and more economical alternative.
An Evaluation of the Lineage of Brucella Isolates in Turkey by a Whole-Genome Single-Nucleotide Polymorphism Analysis
(Mdpi, 2024) Akar, Kadir; Holzer, Katharina; Hoelzle, Ludwig E.; Oz, Gulseren Yildiz; Abdelmegid, Shaimaa; Baklan, Emin Ayhan; Elkhayat, Manar
Brucellosis is a disease that is commonly found in Turkey among both humans and animals. It causes significant economic losses in the livestock industry and is of great public health concern. This study focused on analyzing the genetic makeup of Brucella isolates from Turkey using a core-genome single-nucleotide polymorphism (cgSNP) analysis. This analysis showed potential links between the Turkish isolates and those from neighboring countries and other parts of the world. This highlights the importance of implementing strict measures to control the spread of brucellosis throughout the country. Brucellosis is a disease caused by the Brucella (B.) species. It is a zoonotic disease that affects farm animals and causes economic losses in many countries worldwide. Brucella has the ability to persist in the environment and infect the host at low doses. Thus, it is more important to trace brucellosis outbreaks, identify their sources of infection, and interrupt their transmission. Some countries already have initial data, but most of these data are based on a Multiple-Locus Variable-Number Tandem-Repeat Analysis (MLVA), which is completely unsuitable for studying the Brucella genome. Since brucellosis is an endemic disease in Turkey, this study aimed to examine the genome of Turkish Brucella isolates collected between 2018 and 2020, except for one isolate, which was from 2012. A total of 28 strains of B. melitensis (n = 15) and B. abortus (n = 13) were analyzed using a core-genome single-nucleotide polymorphism (cgSNP) analysis. A potential connection between the Turkish isolates and entries from Sweden, Israel, Syria, Austria, and India for B. melitensis was detected. For B. abortus, there may be potential associations with entries from China. This explains the tight ties found between Brucella strains from neighboring countries and isolates from Turkey. Therefore, it is recommended that strict measures be taken and the possible effects of uncontrolled animal introduction are emphasized.
The First Report of Brucella Melitensis Biovar 2 Strain Isolated From Cattle in Turkey
(German Multidisciplinary Publishing Center - Gmpc, 2023) Akar, Kadir; Yildiz Oz, Gulseren
Bovine brucellosis is an infectious zoonotic disease of great impact on animal welfare and has significant economic implications on livestock farm worldwide. The disease is caused primarily by Brucella abortus (B. abortus), while B. melitensis is less common, and B. suis infection is rare. B. melitensis is the most common causative agent of brucellosis in small ruminants and humans. Although the main host of B. melitensis is considered to be small ruminants, this bacterium is also present in large ruminants. Despite brucellosis has been eradicated in many European countries, it is still endemic in Mediterranean countries and Turkey. The most prevalent Brucella species in the Mediterranean basin and Turkey is B. melitensis biovar (bv) 3. Previous studies have reported that B. melitensis bv2 is quite low in Turkey. This is the first study to isolate B. melitensis bv2 from cattle in Turkey. The strains were characterized using classical biotyping methods and then were molecularly confirmed. Multilocus variable number tandem repeat analysis (MLVA-16) typing of the strains revealed a novel genotype (1-5-3-13-32-3-2-4-41 8 5 4 3 3 7), which matches the Multilocus sequence typing (MLST) profiles in the database of ST8 (3 2 3 2 1 5 3 2 8). These results indicate that B. melitensis bv2 can easily infect cattle and this has to be considered in the epidemiology and control of bovine brucellosis. Circulating the highly pathogenic B. melitensis bv2 in cattle farms is of public health concern.
Genomic Analysis of Brucella Isolates From Animals and Humans, Türkiye, 2010 To 2020
(Eur Centre Dis Prevention & Control, 2024) Akar, Kadir; Brangsch, Hanka; Jamil, Tariq; Oz, Guelseren Yildiz; Baklan, Emin Ayhan; Eroglu, Buket; Wareth, Gamal
Background: Brucellosis is a bacterial zoonosis causing severe illness in humans and animals and leading to economic losses in the livestock production in T & uuml;rkiye and other endemic countries. Aim: We aimed at investigating genomic differences of Brucella isolates from animals and humans in T & uuml;rkiye. Methods: We used whole genome sequencing (WGS) to assess the genetic diversity of Brucella isolates from 41 provinces in T & uuml;rkiye and compared with isolates from other countries. We applied allele-based typing and core genome single nucleotide polymorphism (cgSNP) determination. Results: Of the 106 Turkish Brucella isolates included, 57 were B. abortus and 49 were B. melitensis. . One B. melitensis and two B. abortus isolates were identified as vaccine strains. Most (n = 55) B. abortus isolates clustered in three major branches, with no spatial discernible pattern. Of the B. melitensis isolates, , 48 were assigned to the Eastern Mediterranean lineage with no discernible patterns between host species, location and sampling date. The Turkish isolates clustered with isolates from neighbouring countries such as Greece and Syria, but some also with isolates from human patients in European countries, like Germany, Norway and Sweden, suggesting that the source may be travel- related. Conclusion: Several B. melitensis and B. abortus lineages are circulating in T & uuml;rkiye. To decrease the prevalence and prevent brucellosis in animals and humans, stricter control measures are needed, particularly in areas where humans and animals have close contact. Furthermore, illegal transportation of animals across borders should be more closely controlled and regulated.
Genotyping of Brucella Isolates From Animals and Humans by Multiple-Locus Variable-Number Tandem Repeat Analysis (Mlva)
(Elsevier Sci Ltd, 2023) Ozmen, Murat; Ozgen, Ediz Kagan; Sayi, Orbay; Putur, Elif Karadeniz; Okumus, Biray; Yilmaz, Sibel Iba; Akar, Kadir
This study investigates country-wide genotype variations through the genotyping of Brucella strains isolated from domestic ruminants and humans. The Brucella spp. isolated from samples taken from animals and humans were first identified as B. abortus and B. melitensis by real-time PCR, and the MLVA-16 approach was then used for the genotyping of the identified isolates. For the study, 416 Brucella spp. were isolated from aborted fetus samples examined between 2018 and 2021, and 74 Brucella spp. from infected humans. Of the 74 human isolates analyzed, 1.3% were identified as B. abortus and 98.7% (73/74) as B. melitensis. The MLVA-16 typing method revealed 30 clonal groups for B. abortus and 37 clonal groups for B. melitensis from which the dominant genotypes and similarities with human isolates in Turkiye were determined.
Human Brucellosis and Associated Risk Factors in the Middle East Region: a Comprehensive Systematic Review, Meta-Analysis, and Meta-Regression
(Cell Press, 2024) Dadar, Maryam; Al-Khaza'leh, Ja'far; Fakhri, Yadolah; Akar, Kadir; Ali, Shahzad; Shahali, Youcef
Introduction: Human brucellosis, caused by the bacteria Brucella, , is a significant infectious disease globally, also known as Mediterranean fever or Malta fever. Methods: This meta-analysis and systematic review focuses on the prevalence rate, risk factors, and diagnostic methods of human brucellosis in Middle Eastern countries, where the disease remains a significant public health issue. The analysis included 92 studies conducted between 1993 and 2024, following specified inclusion and exclusion criteria among symptomatic patients across these countries. The prevalence rate was calculated by dividing positive samples by total samples. The I2 index and Chi-squared test assessed study heterogeneity. Significant heterogeneity occurred when I2 exceeded 50 %. Results: The highest prevalence rate was observed in Oman, Lebanon, and Kuwait countries, emphasizing a notable burden of Brucella infection in these regions. Risk factors associated with human brucellosis were identified, with unpasteurized dairy consumption and close animal contact being predominant. Various professions such as farmers, dairy factory workers, and agriculture workers showed higher prevalence rate. Gender analysis indicated a prevalence rate of 18.02 % (95 % CI: 11.55-25.51 %) in males and 13.61 % (95 % CI: 10.8-16.68 %) in females, with no significant difference in hospitalization rates. The prevalence rate of Brucella spp. was varied across detection methods, with immunocapture agglutination assay (Brucellacapt) showing the highest estimated prevalence rate of (44.04 %, 95 % CI: 27.71-61.04), followed by PCR (39.84 %, 95 % CI: 20.14-61.39) and culture (29.22 %, 95 % CI: 17.89-42.03). Among Brucella species, Brucella melitensis (B. melitensis) and Brucella abortus (B. abortus) were the highest prevalence rate. Although statistically insignificant, the meta-analysis also revealed an upward trend in prevalence rate from 1993 to 2024, (P value = 0.277). Discussion: This comprehensive review emphasizes the need for tailored strategies to control brucellosis in the Middle East, highlighting the importance of timely diagnosis, public awareness, and effective treatment protocols. These findings provide valuable insights for policymakers and healthcare professionals working to mitigate the impact of this disease in the region.
Investigation of Brucella Spp. Presence of Amphibians Collected From Some Regions in Türkiye
(2024) Akar, Kadir; Yücetepe, Ayfer Güllü; Öz, Gülseren Yıldız; Baklan, Emin Ayhan; Eroglu, Buket; Atıl, Eray; Erdenlig, Sevil
Brucellosis is a zoonotic disease that causes economic losses in many countries worldwide, especially in livestock. Firstly, the African bullfrog and then B. inopinata and B. microti-like strains have been identified in various frog species worldwide. This study aimed to determine the presence of Brucella in amphibian frogs for the first time in Türkiye by bacteriological and molecular methods. Internal organ samples (spleen, liver, lung, kidney, etc.) of 150 frogs from different regions of Türkiye (Edirne/Ipsala (53 pieces), Adıyaman/Kâhta (97 pieces)) were used for the isolation of Brucella spp. As a result of Brucella genus-specific PCR (Polymerase Chain Reaction) and Multiplex PCR methods performed with these extracts, no positivity was detected in the frog samples taken from Edirne province, while in 4 of the frog samples taken from Adıyaman province, bands of approximately 250, 600, 700, 1000, 1500 and 3000 bp were observed in Multiplex PCR. Sphingomonas paucimobilis (S. paucimobilis) was identified by analysing the culture obtained from these samples with VITEK®2:Healtcare. As a result, the presence of Brucella spp. could not be detected both bacteriologically and molecularly in the study samples. However, observing similar multiple bands in multiplex PCR suggested that this bacterium and Brucella species are phylogenetically close. It was determined that S. paucimobilis, which belongs to the same class as Brucella species under the Alphaproteobacteria, may be dangerous for people who work on farms where frogs are raised for human consumption. This is because the bacteria can cause opportunistic infections, particularly in immunocompromised humans. Therefore, it may be imperative to take the appropriate precautions.
Investigation of the Correlation Between Elisa and Serum Amyloid a in the Diagnosis of Bordetella Bronchiseptica in Dogs
(Wiley, 2025) Akar, Kadir; Golen, Gokcenur Sanioglu; Ekin, Ismail Hakki
BackgroundBordetella bronchiseptica is an essential bacterial pathogen characterized by chronic respiratory disease in dogs known as Kennel cough. The presence of causative antibodies in animals can also be detected by lipopolysaccharide antigen-based enzyme linked immunosorbent assay (ELISA). In recent years, it has been determined that there is a significant relationship between acute phase proteins and diseases, and disease follow-up can be done within the framework of this relationship.MethodsIn this study, blood sera from 150 dogs in an animal shelter in Van province were evaluated for B. bronchiseptica by the homemade ELISA method, and their correlations with serum amyloid A (SAA) were investigated. Blood serum samples were analysed for antibodies against B. bronchiseptica using a homemade ELISA method. Positive animals were also molecularly confirmed using nasal swabs by PCR. A commercial ELISA kit determined SAA levels in blood sera.ResultsEighteen (12%) of the analysed blood serum samples were found positive by the homemade ELISA method. SAA concentrations in the positive blood sera were elevated from 12.7 to <= 38.98 mg/L. SAA concentrations in blood sera serologically positive for B. bronchiseptica were statistically significant.ConclusionsIn this study, in which the relationship between SAA concentration and B. bronchiseptica was investigated for the first time in Turkey, it was concluded that SAA concentration analysis may help diagnose and monitor the disease. In addition, the presence and prevalence of this critical and zoonotic agent causing chronic respiratory tract disease in dogs in Van province was revealed for the first time in this study.
Investigation of the Presence of Mycoplasma Agalactiae in Sheep and Goats in Bitlis Region
(2025) Koçyiğit, Atanur; Akar, Kadir
Bu araştırmada, Bitlis yöresinde süt çekilmesi, sütün azalması ve renk değişikliği gibi klinik semptom gösteren küçük ruminantlarda Mycoplasma (M.) agalactiae varlığının moleküler tekniklerle araştırılması amaçlandı. Bu amaçla Bitlis merkez ve ilçelerinde bulaşıcı agalaksi klinik semptomları gösteren hayvanlardan süt, kan, göz sıvabı, kulak sıvabı ve eklem sıvısından toplanan toplam 120 adet örnek çalışmada kullanıldı. Alınan örneklerin DNA izolasyonları, ticari DNA izolasyon kit protokollerine göre gerçekleştirildi. Daha sonra M. agalactiae tür spesifik olarak belirlenmiş 2 farklı primer çifti (polC ve uvrC) kullanılarak Polimeraz Zincir Reaksiyonu (PCR) aşaması gerçekleştirildi. polC gen bölgesi için süt örneklerinde %44.08, örneklerin tamamında ise %34.16 oranında pozitiflik saptandı. uvrC gen bölgesi için süt örneklerinde %36.55, örneklerinin tamamında ise %28.30 oranında pozitiflik saptandı. Her iki gen pozitif olan örneklerin tamamının keçi süt örneklerinden olduğu belirlendi. Sonuç olarak, her iki gen bölgesi için yapılan analizlerle tespit edilen pozitifliklerin büyük çoğunlukla her iki analiz için de uyumlu olduğunu gözlendi. Çalışma Bitlis bölgesinde M. Agalactiae'nın varlığının araştırıldığı ilk çalışma olma niteliğindedir. Etkenin bölgedeki yaygınlığını belirlemek için periyodik ve daha geniş alana yayılabilecek çalışmalara ihtiyaç bulunmaktadır. Ayrıca bölgede sirküle olan bulaşıcı agalaksi etkeninin kontrol altına alınmasına, eradikasyon stratejileri amacıyla daha ileri düzey moleküler çalışmaların yapılmasının uygun olduğu kanaatine varıldı.
Sığır Fekal Örneklerinden Salmonella Spp. ve Escherichia Coli O157:h7’ye Özgü Bakteriyofaj Varlığının Araştırılması
(2023) Gölen, Gökçenur Sanioğlu; Akar, Kadir
Salmonella Typhimurium (S. Typhimurium) ve Escherichia coli (E. coli) O157:H7, virülanslarına, antimikrobiyal dirençlerine ve yüzeylerde hayatta kalmalarına katkıda bulunan bakteriyel biyofilmler oluşturan ve dünya çapında ciddi gıda zehirlenmelerine neden olan önemli gıda kaynaklı patojenlerdir. Bakteriyofajlar, gıda kaynaklı patojenleri kontrol etmek adına her geçen gün artarak kullanılan antibakteriyel maddeler olup ayrıca antibiyotik direncin gelişmesine karşı çözümde rol oynamaktadır. Ayrıca bakteriyofajlar atık sularda, doğada ve hayvansal atıklarda olabileceği gibi birçok gıda maddesinde bulunmaktadır. Bu çalışmanın amacı sığır altlıklarından alınan dışkı örnekleri kullanılarak ülkemizde ve dünyada sık rastlanan Salmonella türleri. ve E. coli infeksiyonları ile mücadele etmede etkili olabilecek, ülkemizde sirküle olan Salmonella spp. ve E. coli spesifik spot test ile fajların purifikasyonu ve litik etkilerinin belirlenmesi amaçlanmıştır. Bu çalışmada 3 tane S. Typhimurium ve 1 tane E. coli O157:H7 bakteriyofajları izole edilerek, litik aktiviteleri belirlenmiştir. Sonuç olarak bu çalışmada Aksaray ilinden pürifiye edilen Salmonella Typhimurium ve E. coli O157:H7 bakteriyofajlarının tespit edilen litik aktiviteleri ile Salmonella Typhimurium ve E. coli O157:H7 kaynaklı enfeksiyonların tedavisi ve gıda endüstrisinde korunma çalışmalarına ışık tutabileceği düşünülmektedir.
Van‘da Klinik Örneklerde Listeriosis’in Varlığının Araştırılması
(2024) Akar, Kadir; Gölen, Gökçenur Sanioğlu; Babaoglu, Alı Rıza; Ekın, Ismail Hakki
Listeriozis, dünya genelinde insanlar ve çeşitli hayvanlarda abortusa neden olan bir enfeksiyondur. Etken özellikle ruminant gibi çiftlik hayvanlarının dışkılarıyla saçılarak, kontamine silaj ve yemlerin sindirim yoluyla alınmasıyla bulaşan zoonoz karakterli bir özelliğe sahiptir. Bu çalışmada koyunlara ait 120 örnek (79 abort materyali, 41 beyin materyali) kullanılarak Listeria spp. varlığının araştırılması hedeflendi. Bu amaçla Listeria spp. için spesifik oligonükleotid çiftlerinin kullanıldığı polimeraz zincir reaksiyonu (PZR) yöntemi kullanıldı. Yapılan değerlendirmede Van ili ve ilçelerine ait abort materyallerinin tamamı negatif bulundu. Ancak klinik olarak sinirsel semptom gösteren koyunlardan Erciş ve Gevaş ilçelerinden birer tane olmak üzere toplam 2 (%4.87) adet örnekte pozitiflik saptandı. Sonuç olarak abort etkenleri arasında yer alan Listeria spp.’nin Van ilinde sporadik seyrettiği belirlendi. Bu durumun Van il genelinde küçükbaş hayvancılıkta silaj kullanımının oldukça düşük olmasından kaynaklanabileceği kanaatine varıldı. Hastalığın yöredeki seyri için periyodik çalışmaların yapılması gerektiği düşünüldü.