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Öğe Comparative analysis of microscopic and molecular methods for the diagnosis and post-treatment monitoring of Echinococcus granulosus sensu stricto in experimentally infected dogs: Initial findings from digital PCR(Elsevier BV, 2025-07) Muhammed Ahmed Selcuk; Figen Celik; Sami SimsekCystic echinococcosis (CE), caused by Echinococcus granulosus sensu lato, is a significant zoonotic disease with profound public health and economic impacts. This study evaluated egg detection methods, molecular diagnostics, and post-treatment shedding dynamics in experimentally infected dogs. Three three-month-old male dogs were included in the study. Two experimental dogs (ED-1, ED-2) were orally infected with 20,000 E. granulosus protoscoleces, while one served as a control (CD). Stool samples were collected daily over 50 days post-infection and 30 days post-treatment. Detection of E. granulosus eggs was performed using the fülleborn and sieving flotation methods. Genomic DNA was extracted, and molecular analysis was conducted via conventional PCR, Real-time quantitative PCR (qPCR), and digital PCR (dPCR). Egg detection in stool samples began on days 44-46 post-infection using flotation methods, while PCR detected parasite DNA as early as day 20. Both qPCR and dPCR consistently detected parasite DNA from day 1 to day 50 post-infection, with increased sensitivity observed after day 23. Treatment eliminated viable egg shedding within 2-4 days and post-treatment monitoring revealed intermittent detection up to day 30, with dPCR identifying copy numbers even when qPCR Ct values were undetectable. These findings highlight the superior sensitivity of molecular methods in early detection and their persistence in identifying DNA beyond egg-shedding periods. This raises important questions about interpreting molecular positivity in the prepatent phase and its implications for surveillance and diagnostics.Öğe Serodiagnosis of hydatidosis in sheep and goats in Siirt province, Türkiye, and an alternative approach to the interpretation of serology results(Elsevier BV, 2024-02) Burcak Aslan Celik; Muhammed Ahmed Selcuk; Figen Celik; Ozgur Yasar Celik; Muhammet Uslug; Afra Sena Tekin; Kerem Ercan; Sami SimsekCystic echinococcosis (CE) is one of the most important zoonotic diseases worldwide, affecting both humans and animals, caused by Echinococcus granulosus sensu lato. The diagnosis of CE is based on the evaluation of imaging, serological, molecular and clinical findings. The aim of this study was to develop a diagnostic method that can be used for herd-based diagnosis of CE, which is commonly observed in sheep and goats, and to prevent economic losses by treating positive animals. Antigen B rich partially purified cyst fluid antigen was prepared from a sheep liver hydatid cyst. To determine the sensitivity and specificity of the antigen B-rich partially purified cyst fluid antigen, a Western blot test was first performed using hydatid cyst positive sera from 24 sheep and 20 goats collected from the abattoir and sera from 27 newborn lambs and 23 goat kids as negative controls. Subsequently, 500 sera were collected from 250 sheep and 250 goats from Siirt province and its districts. The strongest band was at 72 kDa, followed by bands at 55 kDa and 28 kDa in SDS-PAGE. According to the 43 kDa band as reference the sensitivity was 62.5 %. and specificity was 96.2 % in sheep. Besides, of the 250 sheep sera collected from the field, and the seroprevalence was 52.80 %. In goats, the sensitivity of WB was 55 % and the specificity was 86.9 %. On the other hand, the seroprevalence was 44.4 % in goats. In conclusion, the production of antigens that are easy to prepare, inexpensive and can be used in field screening for CE will be important in terms of treating positive sheep (even when CE and other helminth parasites are present together) with albendazole and preventing economic losses.Öğe Serological diagnosis and post-treatment monitoring of Echinococcus granulosus in experimentally infected dogs using crude and recombinant fibronectin antigens(Elsevier BV, 2025-07) Figen Celik; Muhammed Ahmed Selcuk; Muhammet Uslug; Sami SimsekEchinococcus granulosus is a zoonotic helminth responsible for cystic echinococcosis, a significant public health concern. The diagnosis of E. granulosus infections in definitive hosts, such as dogs, is challenging due to the absence of clinical signs. This study aimed to evaluate the diagnostic performance of crude (EgSCA) and recombinant (rEgFN162) antigens for the detection of E. granulosus infection in dogs using ELISA and Western blot assays. Additionally, it sought to identify the most suitable antigen and method for population-based screening and post-treatment monitoring. Adult E. granulosus parasites were collected from experimentally infected dogs using arecoline hydrobromide purgation. Soluble crude antigen (EgSCA) was prepared through freeze-thaw cycles, sonication, and filtration, while recombinant fibronectin protein (rEgFN162) was obtained via gene cloning, expression, and purification in E. coli. The antigenic properties of EgSCA and rEgFN162 were analyzed by SDS-PAGE and Western blot. ELISA assays were performed to assess IgG and IgM responses in experimentally infected and treated dogs. Based on IgG ELISA results, EgSCA showed a sensitivity of 96.66 % and specificity of 66.66 %, while rEgFN162 demonstrated a sensitivity of 76.66 % and specificity of 46.66 %. In Western blot analysis, EgSCA achieved a sensitivity of 90 % and specificity of 83.33 %, whereas rEgFN162 showed 66.66 % sensitivity and 73.33 % specificity. The recombinant antigen showed a higher ability to differentiate E. granulosus infections from other helminth infections. The findings suggest that rEgFN162 is a promising candidate for the serodiagnosis of E. granulosus in dogs, with potential applications in epidemiological studies and post-treatment follow-up. Further validation with larger sample sizes is needed to confirm its diagnostic accuracy in natural infections.
