Browsing by Author "Çiftçi M.K."

Now showing 1 - 3 of 3

Comparison of Histopathological, Immunohistochemical and Real-Time PCR Methods for Diagnosis of Listeriosis in Ruminants with Encephalitis
(2022-01-01) Hatipoğlu F.; Terzi F.; Özdemir Ö.; Ortatatli M.; Çiftçi M.K.; Ateş M.B.
Encephalitic listeriosis is the most significant purulent encephalitis in ruminants and is a very common endemic problem in sheep, cattle, and goats. In this study, it was aimed to compare the presence of Listeria (L.) monocytogenes revealed by immunohistochemical (IHC) and Real-Time PCR methods with histopathological findings obtained from the archive materials. The study material consisted of pons and medulla oblongata paraffin tissue of 100 ruminants (9 cattle, 4 calves, 44 sheep, 38 lambs, and 5 goats). Positivity was obtained by the IHC method in 46 (46%) and by the Real-Time PCR method in 21 (21%) of 100 cases. In the L. monocytogenesis antigen IHC scoring, more severe staining was observed in sheep and goats (P>0.05). In the IHC positive cases, microabscess was more severe in sheep and goats than in cattle and lambs (P<0.05). In addition, 19 patients had Coenurus cerebralis cysts, and 3 of them were found to be positive for the IHC agent of Listeria. It was concluded that IHC and PCR methods can be used to detect L. monocytogenes from paraffin blocks, but the IHC method is a more effective method than PCR in revealing the presence of antigen from paraffin blocks stored for many years.
Detection of botulinum neurotoxin serotypes c and d, and their effects on expressions of snap-25 and synaptobrevin in ruminants: An immunohistochemical study^[1,2]
(2021-01-01) Ateş M.B.; Terzi F.; Çiftçi M.K.; Ortatatli M.; Çelik Z.
In humans and animals, botulism is a disease characterized by generalized and progressive paralysis caused by Clostridium botulinum neurotoxins (BoNT). BoNTs, defined in seven different antigenic types (A to G), proteolyze SNAREs (synaptosomal-associated protein/SNAP-25 and synaptobrevin) responsible for acetylcholine release in peripheral cholinergic neurons, and thus cause flaccid paralysis and death. Currently, mouse experiments are considered the reference method for definitive diagnosis. However, new diagnostic methods that are fast and accurate and would not raise ethical issues need to be developed. Therefore, using antibodies specific to the toxoid forms of BoNTs, the presence of BoNT-C and/or BoNT-D was investigated by immunohistochemical method (IHC) in the study. The tissues of thirty ruminants (twenty cattle, seven sheep, three goats), which had the clinical and pathological findings of botulism and a herd history of the disease, were used as material. BoNTs were detected with IHC in sixteen of the thirty ruminants as three BoNT-C, eleven BoNT-D, and two BoNT C+D. In the mouse experiments, BoNT was isolated in only three cases (two BoNT-D, one BoNT-C). Additionally, being responsible for the clinical findings of botulism, the interaction of BoNTs with SNAP-25 and synaptobrevin was investigated using IHC. It was determined that BoNT-C specifically reduces the expression of SNAP-25, and BoNT-D reduces the expression of synaptobrevin and partially SNAP-25. It was concluded that additional studies may be valuable to investigate the use of IHC in the diagnosis of botulism.
The usability of cytological and immunocytological methods for rapid diagnosis of encephalitic listeriosis in ruminants
(2021-01-01) Özdemir Ö.; Ortatatli M.; Terzi F.; Hatipoğlu F.; Çiftçi M.K.; Ateş M.B.
Although the clinical and pathological findings are important in the diagnosis of listeriosis, to isolation or to be shown the presence of the bacterium must be required for the definitive diagnosis. This study aims to investigate the availability of imprint cytological (IC) and immunocytochemical (ICC) methods in comparison with histopathological and immunohistochemical (IHC) methods for the rapid diagnosis of encephalitic listeriosis. In the study, the touching and smear preparations taken from the pons and medulla oblongata of 25 ruminants suspected with listeriosis by neurological symptoms were stained with modified giemsa and also with ICC technic for revealing antigens, as a new method. Same tissue sections were stained with Hematoxylin&Eosin and IHC methods too, and examined under light microscope by scoring. In IC examinations, there were intensive neutrophils in 14 cases and few neutrophils in 4 cases, and no neutrophils were observed in 7 cases. In histopathological examinations, 13 of these 14 cases revealed typical microabscesses and listeria positivity in IHC staining. ICC positivity was detected in 12 (92.3%) of the listeria positive 13 cases. A highly positive correlation was observed among cytology (14), ICC (12), histopathology and IHC (13) scores (r2> 0.8; P<0.01). In conclusion, the cytological examination of the pons and medulla oblongata of listeriosis-suspected ruminants revealed that a rapid pre-diagnosis could be made with the presence of intense neutrophils. Also, with ICC staining of cytological preparations, the diagnosis could be performed with 92.3% accuracy. Since ICC is an easy and fast method, it is concluded that it can be used safely especially in field studies, along with cytological examination.