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    A Comparative Pathomorphological Findings Between Leiurus abdullahbayrami and Androctonus crassicauda (Scorpion: Buthidae) Envenomation in Rabbit Animal Model
    (Iranian Scientific Society Medical Entomology, 2019) Ozkan, Ozcan; Alcigir, Mehmet Eray
    Background: The aim of study was to compare macroscopical and histopathological findings between venoms belonging to two scorpion species, Androdoctonus crassicauda, and the newly discovered Leirus abdullahbayrami. Methods: The animals used in this experimental study were fifteen New Zealand bred rabbits. Three groups were constituted as group I (L. abdullahbayrami group, n= 6), group II (A. crassicauda group, n= 6) and group III (control group, n= 3). The animals in the L. abdullahbayrami group and the A. crassicauda group were envenomed through an intravenous route. The rabbits were monitored for the first 24h following the envenomation. The animals dead within that time period were examined and all animals were sacrificed and standard necropsy process was performed at 24h. Results: The pathomorphological findings from group I were found to be more severe than those observed in group II. The venom from the newly identified L. abdullahbayrami has a greater effect than the venom from the A. crassicauda. Moreover, as this was a rabbit modeling study, the L. abdullahbayrami might pose the most serious health threat to infants in particular due to their smaller body weight. Conclusion: These findings will provide a better understanding of envenomation of human beings in terms of the possible consequences of scorpion toxication on the organs.
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    Encephalitozoonosis infection in a traditional rabbit farm with neurological manifestations
    (Elsevier Science Bv, 2018) Ozkan, Ozcan; Alcigir, Mehmet Eray
    Encephalitozoon cuniculi, a zoonotic and opportunistic pathogen, can cause latent infection, especially in lagomorphs. Nowadays, this member of the Eukaryotes has drawn significant attention in the fields of veterinary and public health. The purpose of this study was to determine the seroprevalence of infection in a New Zealand rabbit farm that has a clinical history of neurological manifestations including head tilt ataxia, aggressiveness, seizures, and circling and rotational movements around the body length axis, but the general conditions and food intake were normal. Blood samples were taken from 42 breeding rabbits and researched for E. cuniculi anti-bodies. Out of that, 25 (59%) animals resulted positive against the pathogen. The rabbit was found to be sero-positive for E. cuniculi antibodies, but negative for Toxoplasma gondii and Listeria monocytogenes antibodies. Hematological and serum biochemical parameters were measured at reference intervals. No brain tissue impairment was observed the computed tomography (CT) scan. As a result of these histopathological findings, the brain cortex presented severe neuronal degeneration and partial myelin loss, with reactive diffuse gliosis against the parasite spores was observed to the histopathology. These results are possibly related to the early stage of infection because the parasitic infestation comprise long time spreading. E. cuniculi DNA was detected on brain tissues using polymerase chain reaction (PCR), and it partial DNA sequence was identified as E. cuniculi genotype I.
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    The First Molecular Detection and Genotyping of Encephalitozoon cuniculi in Rabbit's Eye in Turkey
    (Kafkas Univ, Veteriner Fakultesi Dergisi, 2018) Ozkan, Ozcan; Karagoz, Alper; Kocak, Nadir; Alcigir, Mehmet Eray
    Encephalitozoon cuniculi was first recognized as the disease agent in rabbits in 1922. The genotype of E. cuniculi isolated from laboratory rabbits with the neurologic disease was described as genotype I. In the eye, this parasite causes damage to the lens, causing phacoclastic uveitis and cataracts. Intraocular infection often occurs in cases of transplacental transmission. There has been no report on the molecular diagnosis of the parasite in Turkey. The current study is the first report on the detection of E. cuniculi spores using the molecular method in Turkey. In our previous study, a rabbit breeding facility was determined seropositive for E. cuniculi infection monitored for five years in terms of clinical symptoms. An autopsy was performed for a definite diagnosis of the infection. Samples were stained according to the hematoxylin-eosin (H&E) staining after tissue processing procedure and histopathologic analysis was performed. In addition to, the samples for DNA extraction were also taken during the autopsy. ECUNF and ECUNR species-specific primer pairs were used for amplification and genotyping of E. cuniculi. The animals were observed no clinical symptoms except ocular lesion (n=9). Therefore, one of these rabbits was used in the autopsy to definite diagnosis and determination of the damage to the eye. As histopathological, the lesions in the eye were found in the initial or middle stage of progressive infection. The DNA sequence showed that E. cuniculi examined in the present study were genotype I. Possible cause of the visible white mass in the rabbit's eye may be the parasite infection. Therefore, clinicians may consider E. cuniculi as one of the possible causes of ocular lesions in rabbits during daily inspection or ophthalmological examination.
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    Relationship between seropositivity of Encephalitozoon cuniculi and renal biochemical markers in clinically healthy rabbits
    (Ankara Univ Press, 2019) Ozkan, Ozcan; Yucesan, Banucicek; Pekkaya, Selcuk; Alcigir, Mehmet Eray; Gurcan, Ismayil Safa
    Encephalitozoon cuniculi can cause latent disease, especially in lagomorphs and many wild and domestic animals in various countries. This infection is important for veterinary and public health because it is caused by a potentially zoonotic and opportunistic pathogen. The aim of this study was to investigate the relationship between seropositivity of E. cuniculi and renal function markers, which may be useful in predicting the disease in clinically healthy rabbits. In this study, the seropositivity of E. cuniculi infection in clinically healthy rabbits was determined, and necropsy fmdings were evaluated along with the results of renal function tests. In a laboratory rabbit breeding facility, enzyme-linked immunosorbent assay (ELISA) tests showed that 48 (49.5%) of 97 rabbits were seropositive against E. cuniculi. Blood urea nitrogen (BUN) and creatinine levels were significantly elevated in seropositive animals. Two seropositive rabbits were necropsied to confirm the infection. According to histopathological findings in the kidney, degenerative changes and E. cuniculi spores were identified in the tubule epithelia. Serum creatinine (p<0.001) and BUN (p<0.01) levels were found to have a statistically significant relationship with the serological status of rabbits. Serological and histopathological methods are not routinely used in rabbits to diagnose E. cuniculi infection. The kidneys were one of the most affected organs in encephalitozoonosis in rabbits. As revealed in this study, the testing blood urea nitrogen and creatinine levels would be useful for the evaluation of general health status and renal function of the seropositive rabbits, and clinical interpretation as well.
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    Subacute Stage of Encephalitozoon cuniculi Infection in Eye Lesions of Rabbit in Turkey
    (Iranian Scientific Society Medical Entomology, 2018) Ozkan, Ozcan; Alcigir, Mehmet Eray
    Background: Encephalitozoon cuniculi is an opportunistic microsporidian parasite that can affect a number of different species of mammalian animals and humans. The parasite can pose also threat for rabbits even though it causes several sporadic and asymptomatic infections. Infection of eyes is common and clinical symptom of ocular infection may include uveitis and cataracts. We found out subacute findings in naturally infected animals and show here a first described eye lesions as well as central nervous system and kidneys in Turkey. Methods: The rabbits (n: 171) of breeding units were observed to daily clinical examination for infection of E. cuniculi during three years. The eyes of five rabbits (2.9%) showed white intraocular masses or cataracts in the breeding units during daily examinations. The infection was described clinicopathologically in collected organ samples in the animals. During observation, macroscopically, corneal lesions and opacity and impaired lens were taken into attention as well as hyperemia in central nervous system and kidney. Histopathologically, parasitophorous vacuoles pertaining to E. cuniculi were detected in all three tissues during different routine Haematoxylin-Eosin and Gram stainings. Results: Degenerative and necrotic changes in epithelium of cornea and lens and also neurons and tubules were predominantly observed in addition to nonpurulent interstitiel nephritis and encephalitis. Conclusion: The results from study lead to subacute findings especially in eye during natural E. cuniculi infections following asymptomatic and latent changes among breeding colony. The lesions indicated sub-acute stage of E. cuniculi infection in eye lesions of rabbit in Turkey.
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    The effects of antivenom administrations on the brain tissue of experimentally envenomed pregnant rats and their pups with Androctonus crassicauda scorpion venom during organogenesis period
    (Pergamon-Elsevier Science Ltd, 2021) Demirel, Murside Ayse; Alcigir, Mehmet Eray; Ozkan, Ozcan; Turkmen, Merve Biskin
    This study aims to show the changing effects of Androctonus crassicauda venom and A. crasicauda specific antivenom during pregnancy in brain tissue of dams and their pups. Totally, 12 pregnant-Wistar Albino rats were randomly divided into two groups as venom-antivenom administration (n = 6) and control groups (n = 6). In venom-antivenom administration group (VAV), the sublethal dose of A. crassicauda venom dissolved in 1 mL physiological saline solution was subcutaneously (s.c.) injected into pregnant rats during organogenesis period (between 7 and 13 days of pregnancy). Four hours after each venom injection, 1 mL/s.c. dose of the specific antivenom was administered to rats of VAV group. The rats in control group were given sterile saline solution 1 mL/s. c. In both groups, the fetuses were surgically delivered on the 21st day of pregnancy; dams and pups were sacrificed on postnatal 21 days, and their brain tissues were removed. The brain tissue of dams and their pups were evaluated histopathologically and immunohistochemically. To show the neuronal damages, 8-hydroxy-2deoxyguanosine (8-OHDG) and amyloid beta precursor protein (ABPP) immunoexpressions were scored in cerebrum, cerebellum, pons and medulla oblongata of brain. To show the neuroprotection, reelin and beta-arrestin immunoexpressions were scored again in the same way. In this context, 8-OHDG immunoexpressions were increased in neocortex, hippocampus and nucleus accumbens when compared with that of control group. Amyloid beta precursor protein was negative in both groups. Reelin and beta-arrestin partly increased in fore and mid brain of VAV group as a reaction against neuronal damages when compared with that of control pups. The authors believe that prompt intervention using anti-venom to scorpion envenomation can partly stop neuronal damages. This neuroprotection may be increased to high and serial doses of anti-venom to save neonatal lives.
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    The Evaluation of Androctonus crassicauda Antivenom against the Effects of Aegaeobuthus nigrocinctus Scorpion Venom on Autophagy, Apoptosis and Necroptosis
    (Iranian Scientific Society Medical Entomology, 2022) Alcigir, Mehmet Eray; Ozkan, Ozcan
    Background: In this study aimed to show the role of autophagy acting as a seesaw between apoptosis and necroptosis in certain vital organs under the effects of the Aegaeobuthus nigricinctus venom and different dosages of the Androcto-nus crassicauda antivenom administration in mice.Methods: In the venom group (VG), mice (n= 6) were inoculated with 2LD50 A. nigrocinctus venom. In the antivenom administered groups (AVG), the effects of the potency of the A. crassicauda antivenom were evaluated to have a neu-tralization effect against 20LD50 of the A. nigrocinctus venom. After histopathological examination, expressions of mammalian target of rapamycin (mTOR) as an autophagy activator, receptor-interacting serine/threonine-protein kinase 3 (RIPK3) as a necroptosis activator, and caspase-3, caspase-9 as the markers of apoptotic cell death signals were eval-uated by the immunoperoxidase method in addition to DNA in-situ fragmentations by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method.Results: Only in VG, caspases and TUNEL expressions were found to be higher after the envenomation process in contrast to the elevated RIPK3 expressions. mTOR expressions remained almost stable in the organs. In AG, mTOR expressions were further increased in the 30LD50 and 40LD50 groups.Conclusion: There were an increased mTOR expression and stabilized caspases and TUNEL expression in these sub-groups, the RIPK3 expressions were found to be low when compared with all of the antivenom administration groups. Increasing doses of the antivenom drifts more the cells to autophagy while cell fate in organs under envenomation get-ting rid of apoptosis and necroptosis pathways.
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    The lethality, histological, haematological and biochemical alterations in mice envenomated with Aegaeobuthus nigrocinctus venom
    (Pergamon-Elsevier Science Ltd, 2021) Bakir, Fatih; Ozkan, Ozcan; Alcigir, Mehmet Eray; Yagmur, Ersen Aydin
    There is currently no data regarding the toxicity or the in vivo effects of the venom the Aegaeobuthus nigrocinctus species, since it has not been studied thus far according to the best of our knowledge. In the present study, and for the first time, the median lethal dose, the in vivo toxic effects, the histological changes in some of the vital organs were all determined as well as an assessment was made of the histological, biochemical and haematological changes which were caused by the venom injected in mice. The median lethal dose (LD50) of the scorpion venom for mice was found to be 0.38 mg/kg in terms of body weight. The results of the study show that the A. nigrocintus is a potentially lethal scorpion. The evidence related to the venom indicated that it could cause tissue injury in some vital organs. In conclusion, this scorpion venom could cause significant medical complications, and may lead to death, regarding at-risk patients. Therefore, health professionals should be aware of the various scorpion species in their regions and should follow current medical approaches concerning scorpion envenomation.